|
| 1. |
Kizaka-Kondoh, S., Itasaka, S., Zeng, L., Tanaka, S., Zhao, T., Takahashi, Y., Shibuya, K., Hirota, K., Semenza, G.L. and Hiraoka, M.
(2009)
Selective killing of hypoxia-inducible factor-1-actie cells improves survival in a mouse model of invasive and metastatic pancreatic cancer
Clin. Can. Res.
15
,
3433-3441
.
|
| |
Notes:
Hypoxia within solid tumors is a major factor that contributes to cancer progression. Pancreatic cancers express target genes of the transcription factor hypoxia-inducible factor-1. These genes encode proteins involved in angiogenesis, such as vascular endothelial growth factor (VEGF) and other growth factors like insulin-like growth factor (IGF) and proteins associated with the extracellular matrix. The authors of this study investigated whether HIF-1 activity plays a role in progression of pancreatic cancers. They implanted the human pancreatic cancer cells (SUIT-2) that had been transfected with a luciferase reporter in to nude male mice. Mice were either untreated or treated with the prodrug POP33, which appears to increase capsase-3 activity. They sought to determine if POP33 would induce apoptosis in HIF-1 expressing hypoxic cells. Bioluminescent imaging (BLI) was performed in vivo by injecting mice with D-luciferin solution. Caspase activity was also monitored in vivo using a luminescent caspase-3/7 substrate.
(0003997) |
| |
 |
| |
Products: VivoGlo™ Caspase-3/7 Substrate (Z-DEVD-Aminoluciferin) |
| 2. |
Wu, Y., Connors, D., Barber, L., Jayachandra, S., Hanumegowda, U.M. and Adams, S.P.
(2009)
Multiplexed assay panel of cytotoxicity in HK-2 cells for detection of renal proximal tubule injury potential of compounds
Toxicology in Vitro
23
,
1170-1171
.
|
| |
Notes:
The authors describe a multiplexed in vitro assay to detect nephrotoxicity and gain information about mechanism of cell death in HK-2 (human kidney-2) cells. The multiplexed assay involved an LDH assay to detect necrosis, a caspase-3/7 assay to detect apoptosis, a reazurin assay to assess metabolic state, and a DNA dye staining assay to monitor nuclear morphology.
(0004002) |
| |
|
| |
Products: Caspase-Glo® 3/7 Assay | Caspase-Glo® 3/7 Buffer | Caspase-Glo® 3/7 Substrate | CellTiter-Blue® Cell Viability Assay | CellTiter-Glo® Buffer | CellTiter-Glo® Luminescent Cell Viability Assay | CellTiter-Glo® Substrate (lyophilized) | CytoTox-ONE™ Homogeneous Membrane Integrity Assay | CytoTox-ONE™ Homogeneous Membrane Integrity Assay, HTP |
| 3. |
Niles, A.L., Moravec, R.A. and Riss, T.L.
(2009)
In vitro viability and cytotoxicity testing and same-well multi-parametric combinations for high-throughput screening
Current Chemical Genomics
3
,
33-41
.
|
| |
Notes:
The authors review the use of in vitro cytotoxicity testing in drug discovery to characterize the toxic potential of new chemical entities (nce) at the earliest stages of profiling. DOI: 10.2174/1875397300903010033
(0004000) |
| |
|
| |
Products: Apo-ONE® Homogeneous Caspase-3/7 Assay | Apo-ONE® Homogeneous Caspase-3/7 Buffer | Caspase-Glo® 3/7 Assay | CellTiter-Blue® Cell Viability Assay | CellTiter-Fluor™ Cell Viability Assay | CellTiter-Glo® Assay Custom Solution | CellTiter-Glo® Luminescent Cell Viability Assay | MultiTox-Fluor Multiplex Cytotoxicity Assay | MultiTox-Glo Multiplex Cytotoxicity Assay |
| 4. |
Hopkins, M.T., Lampi, Y., Wang, T.W., Liu, Z. and Thompson, J.E.
(2008)
Eukaryotic translation initiation factor 5A is involved in pathogen-induced cell death and development of disease.
Plant Physiol.
148
,
479–489
.
|
| |
Notes:
Genomic DNA from Arabidopsis thaliana was isolated using the Wizard® Genomic DNA Purification Kit. The extracted DNA was used to amplify the 3´ UTR of AteIF5A-2, A. thaliana translation initiation factor 5A, or the entire gene for creating transgenic plants. Leaf discs from wild-type Arabidopsis were infected with Pseudomonas syringae pv tomato DC3000, a virulent strain regulated by AteIF5A-2, using a syringe. After 24 and 72 hours, 0.4cm leaf discs were fixed, labeled using the DeadEnd™ Fluorometric TUNEL System and stained for AteIF5A-2 protein.
(0003979) |
| |
|
| |
Products: DeadEnd™ Fluorometric TUNEL System | Wizard® Genomic DNA Purification Kit | Wizard® Genomic DNA Purification Kit |
| 5. |
Grenier, A.L., Abu-Ihweij, K., Zhang, G., Ruppert, S.M., Boohaker, R., Slepkov, E.R., Pridemore, K., Ren, J.J., Fliegel, L. and Khaled, A.R.
(2008)
Apoptosis-induced alkalinization by the Na+/H+ exchanger isoform 1 is mediated through phosphorylation of amino
Am. J. Physiol. Cell Physiol.
295
,
C883-C896
.
|
| |
Notes:
The authors wanted to examine the role of plasma membrane protein Na+/H+ exchanger isoform 1 (NHE1) in apoptosis. API cells, a NHE1-deficient Chinese hamster ovary cell line, was cotransfected with wild-type NHE1 or mutant NHE1 constructs and destabilized yellow fluorescent protein (YFP). Cells were plated at a density of 1 × 104 cells/well in a 96-well plate with or without FBS. To induce apoptosis in the cells, serum was withdrawn for 24 hours. The ratio of dead-to-live cells was measured using the MultiTox-Fluor Multiplex Cytotoxicity Assay. Cell death was also determined by examining the loss of YFP fluorescence under a microscope.
(0003937) |
| |
|
| |
Products: MultiTox-Fluor Multiplex Cytotoxicity Assay |
| 6. |
Birdsey GM, Dryden NH, Amsellem V, Gebhardt F, Sahnan K, Haskard DO, Dejana E, Mason JC, Randi AM.
(2008)
Transcription factor Erg regulates angiogenesis and endothelial apoptosis through VE-cadherin.
Blood
111
,
33498-33506
.
|
| |
Notes:
These authors showed that the ETS transcription factor Erg interacts with the VE-cadherin promoter region and regulates endothelial apoptosis through this interaction. They demonstrated that inhibition of Erg by siRNA resulted in decreased VE-cadherin mRNA and protein levels, and showed that Erg interacts with the VE-cadherin promoter using a CHIP assay. To show the functional relevance of this interaction, HeLa cells were transfected with a pGL4 Vector containing the VE-cadherin promoter region and an expression vector containing Erg2 cDNA. In this reporter assay, Erg overexpression resulted in ~1.8 fold transactivation of VE-cadherin promoter activity, as measured using the Dual-Luciferase® Reporter Assay System. Inhibition of Erg in human umbilical vein endothelial cells also resulted in a loss of viability and an increase in activation of caspase 3 and caspase 7. The authors showed that apoptosis could be significantly decreased by overexpression of VE-cadherin, indicating that Erg regulates survival partially via its interaction with VE-cadherin. The Caspase-Glo® 3/7 Assay was used to measure caspase activity in these experiments.
(0003872) |
| |
|
| |
Products: Caspase-Glo® 3/7 Assay | Dual-Luciferase® Reporter 1000 Assay System | Dual-Luciferase® Reporter Assay System | Dual-Luciferase® Reporter Assay System 10-Pack |
| 7. |
Cavallaro, M., Mariani, J., Lancini, C., Latorre, E., Caccia, R., Gullo, F., Valotta, M., DeBiasi, S., Spinardi, L., Ronchi, A., Wanke, E., Brunelli, S., Favaro, r., Ottolenghi, S. and Nicolis, S.K.
(2008)
Impaired generation of mature neurons by neural stem cells from hypomorphic Sox2 mutants
Development
135
,
541-557
.
|
| |
Notes:
The authors of this paper investigated the role of Sox2 in neuronal differentiation. Neurospheres were derived from the brains of normal and Sox2 hyphomorphic mice and used to generate differentiated neural cells. In astroglia from cultures containing a Sox2-GFP-expressing lentivirus, ectopic expression of Sox2 correlated with reduced GFAP expression. The authors investigated the role of Sox2 by amplifying binding sites upstream of the GFAP promoter and cloning them upstream of the thymidine kinase promoter in the pRL-TK vector. The Dual-Luciferase Assay System was used to analyze the effect of Sox2 expression on the luciferase reporter gene. The DeadEnd™ Fluorometric TUNEL System was also used to assess apoptosis in some of the neurosphere-derived cultures.
(0003953) |
| |
|
| |
Products: DeadEnd™ Fluorometric TUNEL System | Dual-Luciferase® Reporter 1000 Assay System | Dual-Luciferase® Reporter Assay System | pRL-TK Vector |
| 8. |
Hahn, C.K., Ross, K.N., Warrington, I.M., Mazitschek, R., Kanegai, C.M., Wright, R.D., Kung, A.L., Golub, T.R. and Stegmaier, K.
(2008)
Expression-based screening identifies the combination of histone deacetylase inhibitors and retinoids for neuroblastoma differentiation.
Proc. Natl. Acad. Sci. USA
105
,
9751-9756
.
|
| |
Notes:
The authors designed a high-throughput gene-expression screen to identify compounds that induce a neuroblastoma gene signature in BE(2)-C cells. The screen used Valproic Acid (VPA), an inhibitor of histone deacetylase, as an enhancer. The top hit from the screen was all-trans retinoic acid (ATRA). The authors proposed that ATRA + VPA would reduce cell viability and might induce apoptosis by inducing genes associated with terminal differentiation. They used the CellTiter-Glo® Luminescent Cell Viability Assay in a 96-well format to assess the effects of ATRA and a variety of inhibitors of histone deacetylase on BE(2)-C cell viability.
(0003931) |
| |
|
| |
Products: CellTiter-Glo® Luminescent Cell Viability Assay |
| 9. |
Chang, Q,. Bhatia, D., Zhang, Y., Meighan, T., Castranova, V., Shi, X. and Chen, F.
(2008)
Incorporation of an internal ribosome entry site-dependent mechanism in arsenic-induced GADD45 alpha expression.
Cancer Res.
2007
,
6146–54
.
|
| |
Notes:
Trivalent arsenic (As3+) induces expression of growth arrest- and DNA damage-induced gene 45α (GADD45α), which interacts with intracellular signaling molecules involved in cell cycle regulation, apoptosis and immune response. To characterize GADD45α mRNA expression patterns, total RNA was isolated from untreated (growing) and As3+-treated (arrested) BEAS-2B cells, and GADD45α mRNA was amplified using the AccessQuick™ RT-PCR System. GADD45α mRNA levels were undetectable in growing cells but increased in a time-dependent manner in arrested cells. Reverse transcription was carried out at 45°C for 50 minutes, followed by 35 cycles of PCR.
(0003766) |
| |
|
| |
Products: AccessQuick™ RT-PCR System |
| 10. |
Treeck, O., Pfeiler ,G., Horn, F., Federhofer, B., Houlihan, H., Vollmer, A., and Ortmann, O.
(2007)
Novel estrogen receptor beta transcript variants identified in human breast cancer cells affect cell growth and apoptosis of COS-1 cells.
Mol. Cell. Endocrinol.
264
,
50-60
.
|
| |
Notes:
This study identified two novel transcript variants of the estrogen receptor ERβ that were expressed in the ERα-negative breast cancer cell line MDA-MD-231. These variants were identified after amplification of ERβ transcripts from the breast cancer cell line by RT-PCR. The amplification products were then excised from gels and subcloned into the pTARGET™ Mammalian Expression Vector prior to sequencing. COS1 cells, which do not express the estrogen receptor, were then stably transfected with full-length ERβ or one of the splice variants and the effects on cell proliferation, apoptosis, and estrogen response were evaluated. In COS1 cells expressing either ERβ or the transcript variants cell proliferation decreased and basal apoptosis (caspase 3/7 activity) increased, compared to cells transfected with vector alone. Exposure to therapeutic doses of tamoxifen induced apoptosis in cells expressing the full-length ERβ but not in cells expressing either of the variant isoforms.
(0003618) |
| |
|
| |
Products: Caspase-Glo® 3/7 Assay | CellTiter-Blue® Cell Viability Assay | ImProm-II™ Reverse Transcriptase | M-MLV Reverse Transcriptase Buffer Pack | M-MLV Reverse Transcriptase, RNase H Minus | pTARGET™ Mammalian Expression Vector System |
| 11. |
Lynch, R.A., Etchin, J., Battle, T.E. and Frank, D.A.
(2007)
A small-molecule enhancer of signal transducer and activator of transcription 1 transcriptional activity accentuates the antiproliferative effects of IFN-γ in human cancer cells
Cancer Res.
67
,
1254-1261
.
|
| |
Notes:
STAT1 is a transcription factor that is involved in a variety of cellular processes and behaves like a tumor suppressor in many ways. It is associated with apoptosis, inhibition of cyclin-dependent kinases, and it may mediate the antitumor effects of IFN-γ. The authors of this study designed a bioluminescent reporter assay to identify small molecules that enhance STAT1-dependent gene expression. The bioluminescent assay was performed in 384-well plates using both stably and transiently transfected cell lines. The screening assay was robust, producing a Z´factor value of 0.92, and it identified three compounds that specifically enhanced STAT1-dependent transcriptional activity. Firefly luciferase activity in stable cell lines was assessed using the Bright-Glo® Luciferase Assay System; luciferase activity in the transiently transfected cells was monitored using the Dual-Luciferase® Assay System. Viability of cells in culture was monitored using the CellTiter-Glo® Assay.
(0003732) |
| |
|
| |
Products: Bright-Glo™ Luciferase Assay System | CellTiter-Glo® Luminescent Cell Viability Assay | Dual-Luciferase® Reporter Assay System | pRL-TK Vector |
| 12. |
Wu, J., Apontes, P., Song, L., Liang, P., Yang, L. and Li, F.
(2007)
Molecular mechanism of upregulation of survivin transcription by the AT-rich DNA-binding ligand, Hoechst33342: evidence for survivin involvement in drug resistance.
Nucleic Acids Res.
35
,
2390–2402
.
|
| |
Notes:
To study how Hoechst33342 upregulates the expression and promoter activity of survivin, a novel member of the inhibitor of apoptosis (IAP) protein family, nested deletions of the survivin promoter driving a firefly luciferase reporter gene (pLuc-1430c ) were created using the Erase-a-Base® System. The vector was digested with SalI, the ends filled in using α-phosphorothioate dNTPs, digested a second time with BamHI and subjected to Exonuclease III digestion at 25°C. Aliquots of the 5’ end deletions were removed every 15–30 seconds, religated, transformed and analyzed by PCR and sequencing. Transient transfection experiments were carried out using HeLa cells seeded in 24-well plates and cotransfected 490ng of a pLuc-survivin construct and 10ng of pRL-TK Vector or in U937 cells using 2µg of survivin promoter constructs. After 24 hours, the HeLa cells were treated with Hoechst33342 and harvested 8–24 hours later. For U937 cells, the medium was changed with or without added drugs and the cells lysed after 36 hours. Reporter expression was assessed using the Dual-Luciferase® Reporter Assay System.
(0003697) |
| |
|
| |
Products: Dual-Luciferase® Reporter 1000 Assay System | Dual-Luciferase® Reporter Assay System | Dual-Luciferase® Reporter Assay System 10-Pack | Erase-a-Base® System (minus vectors & bacterial strain) | pRL-TK Vector |
| 13. |
Fan, F. and Wood, K.V.
(2007)
Bioluminescent assays for high-throughput screening
Assay Drug Dev. Technol.
5
,
127–136
.
|
| |
Notes:
The authors of this paper review bioluminescent assay technologies, discussing HTS reporter, cell-based and luciferase biosensor assays. They divide luminescent assays into three basic categories: assays that measure ATP concentration (cell viability and kinase assays), assays that measure changes in luciferase levels (reporter assays, GPCR assays), and assays that measure changes in luciferin levels (protease [including caspase], P450 and MAO assays).
(0003737) |
| |
|
| |
Products: Renilla Luciferase Assay System | BacTiter-Glo™ Microbial Cell Viability Assay | Bright-Glo™ Luciferase Assay System | Calpain-Glo™ Protease Assay | cAMP-Glo™ Assay | Caspase-Glo® 2 Assay | Caspase-Glo® 3/7 Assay | Caspase-Glo® 6 Assay | Caspase-Glo® 8 Assay | Caspase-Glo® 9 Assay | CellTiter-Glo® Luminescent Cell Viability Assay | Dual-Glo® Luciferase Assay System | Dual-Luciferase® Reporter 1000 Assay System | Dual-Luciferase® Reporter Assay System | Dual-Luciferase® Reporter Assay System 10-Pack | Kinase-Glo® Lumine |
| 14. |
Straszewshi-Chavez, A., Visintin, I.P., Karassina, N., Los, G., Liston, P., Halaban, R., Fadiel, A. and Mor, G.
(2007)
XAF1 mediates tumor necrosis factor-α-induced apoptosis and X-linked inhibitor of apoptosis cleavage by acting through the mitochondrial pathway
Journal of Biological Chemistry
282
,
13059-13072
.
|
| |
Notes:
The authors sought to determine the mechanism by which first-trimester trophoblasts resist FAS ligand-induced apoptosis but remain sensitive to TNFα-mediated apoptosis. First trimester trophoblasts express XAF1 [X-linked inhibitor of apoptosis (XIAP)-associated factor 1], which may be involved in regulating their response to proapoptotic signals. The authors created HaloTag™-XAF1 fusion constructs and transiently transfected the first trimester trophoblast cell line (3A). Cells were labeled with the HaloTag™ TMR ligand, and XAF1 was shown to localize to the cytoplasm. 3A cells transiently transfected with the fusion construct were also separated into cytoplasmic and mitochondrial fractions. The fractions were labeled with HaloTag™ TMR ligand. Expression of the fusion peaked at 48 hours after transfection in both mitochondrial and cytoplasmic fractions. TNFα-treatment of 3A cells induced translocation of endogenous XAF1 to the mitochondria. The authors used the Caspase-Glo® Assays to demonstrate activation of caspase-3 and caspase-9 in response to expression of XAF-1. They also show that caspase-3 activation and XIAP cleavage correlate with translocation of endogenous XAF1 to mitochondria. Viability of 3A and primary trophoblasts over expressing XAF1 was evaluated using the CellTiter® 96 AQueous One-Solution Assay.
(0003760) |
| |
|
| |
Products: Caspase-Glo® 3/7 Assay | Caspase-Glo® 9 Assay | CellTiter 96® AQueous One Solution Cell Proliferation Assay | HaloTag® pHT2 Vector | HaloTag® TMR Ligand |
| 15. |
Muraoka-Cook, R.S., Caskey, L.S., Sandahl, M.A., Hunter, D.M., Husted, C., Strunk, K.E., Sartor, C.I., Rearick, W.A., McCall, W., Sgagias, M.K., Cowan, K.H., and Earp, H. S.
(2007)
Heregulin-dependent delay in mitotic progression requires HER4 and BRCA1.
Mol. Cell. Biol.
26
,
6412-6424
.
|
| |
Notes:
Heregulin is a growth factor that binds to the HER/ErbB family of receptors, which includes four HER members. HER1 and HER2 overexpression in breast cancer cells is associated with higher malignancy and poorer prognosis. However, HER4 expression is associated with longer survival and more positive prognosis. The authors of this study investigated HER4-dependent growth inhibition that is mediated by heregulin. The authors determined the absolute levels of HER4 mRNA in several human breast cancer and mouse cancer cell lines. For this determination, relative cell numbers were determined using a CellTiter® AQueous MTS assay. Apoptosis in heregulin-treated or untreated cells was also determined using the DeadEnd™ Colorimetric TUNEL System. The authors concluded that heregulin decreases growth of HER4-positive breast cancer cells, and that this growth inhibition is dependent on BRCA1.
(0003596) |
| |
|
| |
Products: CellTiter 96® AQueous MTS Reagent Powder | CellTiter 96® AQueous One Solution Cell Proliferation Assay | DeadEnd™ Colorimetric TUNEL System | Olomoucine (cdc2 Protein Kinase Inhibitor) |
| 16. |
Budinger, G.R.S., Mutlu, G.M., Eisenbart, J., Fuller, A.C., Bellmeyer, A.A., Baker, C.M., Wilson, M., Ridge, K., Barrett, T.A., Lee, V.Y. and Chandel, N.S.
(2006)
Proapoptotic Bid is required for pulmonary fibrosis.
Proc. Natl. Acad. Sci. U S A
103
,
4604-4609
.
|
| |
Notes:
The authors of this study investigated the role of the mitochondrial-dependent cell death pathway in the development of pulmonary fibrosis after lung injury in a mouse model. The TGFβ1 Emax ImmunoAssay System was used to measure TGFβ1 levels in brochoalveolar lavage fluid from Bid-/- and wildtype mice 5 days after lung injury induced by bleomycin treatment. Mice lacking Bid were protected from pulmonary fibrosis, even although they had similar levels of lung injury, inflammation, and TGFβ1 as wildtype mice 5 days after bleomycin administration. These results indicated that lack of Bid protected against TGFβ1-induced cell death.
(0003471) |
| |
|
| |
Products: TGFβ1 Emax® ImmunoAssay System |
| 17. |
Basgupta, P., Kinkade, R., Joshi, B., DeCook, C., Haura, E. and Chellappan, S.
(2006)
Nicotine inhibits apoptosis induced by chemotherapeutic drugs by up-regulating XIAP and survivin.
Proc. Natl. Acad. Sci. USA
103
,
6332-7
.
|
| |
Notes:
RT-PCR was performed to map the subtypes of nicotinic acetylcholine receptors on A549 cells. cDNA was synthesized using the Reverse Transcription System. Northern blotting to assess XIAP and survivin expression was peformed, and probes were labeled using the Prime-A-Gene® Labeling System. Apoptosis was assessed in nicotine-stimulated cells using a DeadEnd™ TUNEL Assay.
(0003382) |
| |
|
| |
Products: DeadEnd™ Colorimetric TUNEL System | DeadEnd™ Fluorometric TUNEL System | Prime-a-Gene® Labeling System | Reverse Transcription System |
| 18. |
Harada, C., Nakamura, K., Namekata, K., Okumura, A., Mitamura, Y., Iizuka, Y., Kashiwagi, K., Yoshida, K., Ohno, S., Matsuzawa, A., Tanaka, K., Ichijo, H. and Harada, T.
(2006)
Role of apoptosis signal-regulating kinase 1 in stress-induced neural cell apoptosis in vivo.
American Journal of Pathology
168
,
261-269
.
|
| |
Notes:
The authors of this study investigated the role of apoptosis signal-regulating kinase 1 (ASK1) in neural cell apoptosis during retinal development and ischemic injury. Nucleotides 283 to 713 of the ASK1 cDNA were amplified by PCR and cloned into the pGEM®-T Easy Vector, and sense and antisense probes for in situ hybridization experiments were generated. Anti-ACTIVE® p38 polyclonal antibody was used for immunohistochemistry analyses to investigate the localization of phosphorylated p38 in mouse retina.
(0003530) |
| |
|
| |
Products: Anti-ACTIVE® p38 pAb, Rabbit, (pTGpY) | pGEM®-T Easy Vector System I | pGEM®-T Easy Vector System II |
| 19. |
Slot, K.A., Voorendt, M., de Boer-Brouwer, M., van Vugt, H.H. and Teerds, K.J.
(2006)
Estrous cycle dependent changes in expression and distribution of Fas, Fas ligand, Bcl-2, Bax, and pro- and active caspase-3 in the rat ovary.
J. Endocrinol.
188
,
179–192
.
|
| |
Notes:
The authors examined the locations and levels of apoptosis-related Fas, Fas ligand, Bcl-2, Bax and caspase-3 proteins in ovarian tissue throughout the rat estrus cycle. Protein levels were determined using Western blotting, and proteins were localized by immunhistochemistry. The presence of Fas, Fas ligand, Bcl-2, Bax and caspase-3 mRNA in various ovarian tissues was monitored by RT-PCR. Reverse transcription was performed using the ImProm-II™ Reverse Transcription System, 1µg of RQ1 RNase-free DNase-treated RNA and an oligo(dT) primer. One microliter of the reverse transcription reaction was amplified by PCR, and 10µl of the amplification products were analyzed by agarose gel electrophoresis.
(0003724) |
| |
|
| |
Products: ImProm-II™ Reverse Transcription System | RQ1 RNase-Free DNase |
| 20. |
Budhram-Mahadeo, V.S., Bowen, S., Lee, S., Perez-Sanchez, C., Ensor, E., Morris, P.J. and Latchman, D.S.
(2006)
Brn-3b enhances the pro-apoptotic effects of p53 but not its induction of cell cycle arrest by cooperating in trans-activation of bax expression.
Nucleic Acids Res.
34
,
6640–6652
.
|
| |
Notes:
Previously, the POU domain of Brn-3a was shown to interact with p53 and increase cell survival. In this article, the authors explored the possibility that Brn-3b, which shares a POU domain 95% identical to Brn-3a, may interact with p53 and affect its role in apoptosis. To test the protein:protein interaction, GST-Brn-3b fusion protein was bound to glutathione Sepharose beads and incubated with 35S-methionine labeled full-length or truncated p53, prepared using the TNT® T7 rabbit reticulocyte lysate. The luciferase control included in the kit was used as the noninteracting protein control. After washing, the bound proteins were resolved by 12% SDS-PAGE, and the bands examined by radiography. To examine the effect of Brn-3b on two p53-regulated genes, Bax and p21cip1/waf1, ND7 cells were transiently transfected with empty vector, Brn-3b, Brn-3a or p53, or Brn-3 with p53. The reporter gene cotransfected was under the control of wildtype Bax, wildtype p21cip1/waf1, or mutant Bax. A control vector (Renilla luciferase driven by the thymidine kinase promoter) was used for normalization. Forty-eight hours posttransfection, the cells were harvested and reporter levels assessed using the Dual-Luciferase® Reporter Assay System.
(0003597) |
| |
|
| |
Products: Dual-Luciferase® Reporter 1000 Assay System | Dual-Luciferase® Reporter Assay System | Dual-Luciferase® Reporter Assay System 10-Pack | pGL4.74[hRluc/TK] Vector | pRL-TK Vector | TNT® T7 Coupled Reticulocyte Lysate System | TNT® T7 Coupled Reticulocyte Lysate System, Trial Size | TNT® T7 Quick Coupled Transcription/Translation System | TNT® T7 Quick Coupled Transcription/Translation System, Trial Size |
| 21. |
Seehuus, S-C., Norberg, K., Gimsa, U., Krekling, T. and Amdam, G.V.
(2006)
Reproductive protein protects functionally sterile honey bee workers from oxidative stress
Proc. Natl. Acad. Sci. U S A
103
,
962-967
.
|
| |
Notes:
The authors of this paper demonstrate that the pathways controlling lifespan and senescence are linked to the pathways controlling fertility in the honey bee. Using RNAi experiments, the authors demonstrate that vitellogenin protects honey bee workers from oxidative stress. Double-stranded RNA was synthesized from the vitellogenin clone AP4a5 using the RiboMax® T7 System. In additional experiments the authors assessed apoptosis in brain tissue sections using the DeadEND™ Fluorometric TUNEL system.
(0003636) |
| |
|
| |
Products: DeadEnd™ Fluorometric TUNEL System | RiboMAX™ Large Scale RNA Production System—T7 |
| 22. |
Yamauchi, H., Hotta, Y., Konishi, M., Miyake, A., Kawahara, A. and Itoh, N.
(2006)
Fgf21 is essential for haematopoiesis in zebrafish.
EMBO Rep.
7
,
649-54
.
|
| |
Notes:
In this study, a zebrafish homolog of the human Fgf21 gene was identified, and the role of Fgf21 signaling was investigated using Fgf21-knockdown zebrafish embryos. The Fgf21-knockdown embryos lacked erythroid and myeloid cells, but not lymphoid cells or blood vessels. Fgf21 was therefore shown to be required for erythropoiesis, but not for vasculogenesis. Lack of Fgf21 did not appear to affect apoptosis or cell proliferation rates in the intermediate cell mass. The DeadEnd™ Colorimetric TUNEL System was used for apoptosis determination.
(0003582) |
| |
|
| |
Products: DeadEnd™ Colorimetric TUNEL System |
| 23. |
Geiger, G.A, Parker S.E., Beothy, A.P., Tucker, J.A., Mullins, M.C., and Kao, G.D.
(2006)
Zebrafish as a "biosensor"? Effects of ionizing radiation and amifostine on embryonic viability and development.
Cancer Res.
66
,
8172-81
.
|
| |
Notes:
In this study, the Caspase-Glo® 8 and Caspase-Glo® 9 Assays were used to assess caspase activation in Zebrafish embryos after radiation exposure. After irradiation, caspase activation, morphological abnormalities, and DNA fragmentation were observed, all three of which could be partially reversed by treatment with the radiomodifier amifostine. The Caspase-Glo® 9 Assay was shown to be sensitive enough to detect the effects of radiation on as few as 2 embryos. The authors concluded that the Caspase-Glo® Assays provided an effective and convenient means for rapidly assessing the lethal effects of radiation on Zebrafish embryos, and for assaying the ability of the radiomodifier to counteract these effects. The assay could be performed within hours of radiation exposure directly on the embryos in a 96-well plate format.
(0003571) |
| |
|
| |
Products: Caspase-Glo® 8 Assay | Caspase-Glo® 9 Assay |
| 24. |
Patel, V., Longacre, A., Hsiao, K., Fan, H., Meng, F., Mitchell, J.E., Rauch, J., Ucker, D.S. and Levine, J.S.
(2006)
Apoptotic cells, at all stages of the death process, trigger characteristic signaling events that are divergent from and dominant over those triggered by necrotic cells.
J. Biol. Chem.
281
,
4663-4670
.
|
| |
Notes:
Serum-starved, bone marrow-derived macrophages were exposed to apoptotic or necrotic cells, or latex beads (control), and phosphorylation of a variety of signaling molecules was investigated by Western blot. Anti-ACTIVE® MAPK pAb, Anti-ACTIVE® p38 pAb, Anti-ACTIVE® JNK pAb and Anti-ERK 1/2 pAb were used.
(0003428) |
| |
|
| |
Products: Anti-ACTIVE® JNK pAb, Rabbit, (pTPpY) | Anti-ACTIVE® MAPK pAb, Rabbit, (pTEpY) | Anti-ACTIVE® p38 pAb, Rabbit, (pTGpY) | Anti-ERK 1/2 pAb, Rabbit |
| 25. |
Daido, S., Yamamoto, A., Fujiwara, K., Sawaya, R., Kondo, S. and Kondo, Y.
(2005)
Inhibition of the DNA-Dependent Protein Kinase catalytic subunit radiosensitizes malignant glioma cells by inducing autophagy.
Cancer Res.
65
,
4368-4375
.
|
| |
Notes:
The SignaTECT® DNA-Dependent Protein Kinase (DNA-PK) Assay System was used to assess DNA-PK activity in several human malignant glioma cell lines.
(0003409) |
| |
|
| |
Products: SignaTECT® DNA-Dependent Protein Kinase Assay System |