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J. Biol. Chem. 277, 40919-29. Localization versus function of Rab3 proteins. Evidence for a common regulatory role in controlling fusion. 2002

Schlüter, O.M., Khvotchev, M., Jahn, R., and Südhof, T.C.

Notes: These authors used the FuGENE® 6 reagent to transiently transfect PC12 (rat adrenal gland) cells with 2µg DNA at a 3:1 transfection reagent:DNA ratio. Cells were grown to 50-60% confluence in 6-well plates prior to transfection. (4390)

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Infect. Immun. 70, 4571-80. Opsonization modulates Rac-1 activation during cell entry by Leishmania amazonensis. 2002

Morehead, J., Coppens, I., and Andrews, N.W.

Notes: These authors used the FuGENE® 6 reagent to transiently transfect CHO cells with DNA at a 3:1 transfection reagent:DNA ratio. (4396)

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J. Biol. Chem. 277, 26217-24. Transforming growth factor-beta induces expression of receptor activator of NF-kappa B ligand in vascular endothelial cells derived from bone. 2002

Ishida, A., Fujita, N., Kitazawa, R., and Tsuruo, T.

Notes: These authors used the FuGENE® 6 reagent to transiently transfect ST2 mouse bone marrow stroma cells with 1µg DNA at a 3:1 transfection reagent:DNA ratio. Cells were grown to 50-80% confluence in 6-well plates prior to transfection. (4389)

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J. Biol. Chem. 276, 39892-7. IGF-I receptor-induced cell-cell adhesion of MCF-7 breast cancer cells requires the expression of junction protein ZO-1. 2001

Mauro, L., Bartucci, M., Morelli, C., Andò, S., and Surmacz, E.

Notes: These authors used the FuGENE® 6 reagent to transiently transfect MCF7 human breast carcinoma cells with DNA at a 3:1 transfection reagent:DNA ratio. (4393)

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Mol. Cell. Biol. 21, 6626-6639. Silencing of Wnt signaling and activation of multiple metabolic pathways in response to thyroid hormone-stimulated cell proliferation. 2001

Miller, L.D., Park, K.S., Guo, Q.M., Alkharouf, N.W., Malek, R.L., Lee, N.H., Liu, E.T., and Cheng, S.Y.

Notes: In this study, FuGENE® 6 Reagent was used to transiently transfect GC (rat pituitary tumor) cells with plasmid DNA. Cells were plated at 3.5 x 10e5 cells/dish in 35mm dishes and transfected with 1.5µg DNA at a 2.5:1 FuGENE:DNA ratio. (4366)

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J. Virol. 75, 9142–55. Induction of p53-independent apoptosis by simian virus 40 small t antigen. 2001

Gjoerup, O. Zaveri, D. and Roberts, T.M.

Notes: Human osterosarcoma cells were stably transfected using FuGENE and puromycin selection. Transfections were performed in 100mm dishes using 5μg of DNA and a 3:1 ratio of reagent to DNA. (4292)

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J. Biol. Chem. 276, 47038-45. Oncostatin M regulates the synthesis and turnover of gp130, leukemia inhibitory factor receptor alpha, and oncostatin M receptor beta by distinct mechanisms. 2001

Blanchard, F., Wang, Y., Kinzie, E., Duplomb, L., Godard, A., and Baumann, H.

Notes: These authors used the FuGENE® 6 reagent to transiently transfect MCF-7 human breast carcinoma cells and NIH-3T3 mouse embryonic fibroblast cells with DNA at a 6:4 transfection reagent:DNA ratio. (4388)

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Mol. Cell. Biol. 20, 6600-11. A calcineurin-NFATc3-dependent pathway regulates skeletal muscle differentiation and slow myosin heavy-chain expression. 2000

Delling, U., Tureckova, J., Lim, H.W., De Windt, L.J., Rotwein, P., and Molkentin, J.D.

Notes: These authors used the FuGENE® 6 Reagent to transiently transfect 10T1/2 mouse embryonic fibroblasts with 2µg plasmid DNA. Cells were grown in 60mm dishes. (4367)

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J. Biochem. 275, 19250–7. A role for focal adhesion kinase in phenylephrine-induced hypertrophy of rat ventricular cardiomyocytes. 2000

Taylor, J.M., Rovin, J.D. and Parsons, J.T.

Notes: Ventricular cardiomyocytes were isolated from neonatal rats. Cells were plated on 20mm chamber slides precoated with ECM or gelatin. For transfection experiments cells were transfected with an expression vector (3µg DNA) using FuGENE® 6 reagent in serum-free medium. (4275)

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Mol. Cell. Biol. 20, 1982-92. An AU-rich sequence element (UUUN[A/U]U) downstream of the edited C in apolipoprotein B mRNA is a high-affinity binding site for Apobec-1: binding of Apobec-1 to this motif in the 3' untranslated region of c-myc increases mRNA stability. 2000

Anant, S., and Davidson, N.O.

Notes: In this study, FuGENE® 6 was used to transiently transfect 3T3 F442A cells (mouse adherent preadipocytes) with 3µg plasmid DNA. 5 x 10e4 cells were plated in 35mm dishes.  (4568)

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J. Biol. Chem. 275, 12298-305. Change of specificity mutations in androgen-selective enhancers. Evidence for a role of differential DNA binding by the androgen receptor. 2000

Verrijdt, G., Schoenmakers, E., Haelens, A., Peeters, B., Verhoeven, G., Rombauts, W., and Claessens, F.

Notes: These authors used the FuGENE® 6 reagent to transiently transfect HeLa cells with 0.5µg DNA. Cells were cells plated at 6 x 10e4 cells/well 1 day prior to transfection in 24-well plates. (4395)

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J. Biol. Chem. 275, 23661–5. Elevation of mitochondrial calcium by ryanodine-sensitive calcium-induced calcium release. 2000

Nassar, A. and Simpson, A.W.M.

Notes: A10 cells, derived from rat embryonic thoracic aorta, were grown on 16mm coverslips in a 12-well plate and transfected with a plasmid expressing mitochondrially targeted apoaequorin using FuGENE® 6 transfection reagent. The FuGENE® 6 reagent was diluted to 100µl with sterile Dulbecco’s modified Eagle’s medium containing 1% antibiotic and no serum, the diluted reagent was combined with plasmid DNA and 1.5µl of the mixture containing 0.5µg of plasmid DNA was added to each well. (4272)

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J. Biol. Chem. 275, 31162–70. Hepatocyte nuclear factor-3 homologue 1 (HFH-1) represses transcription of smooth muscle-specific genes. 2000

Hoggatt, A.M. et al.

Notes: Rat A10 vascular smooth muscle cells were seeded at 1.4 × 105 cells/dish in 35mm dishes and transfected using FuGENE® 6 transfection reagent. The authors used a total of 2µg of plasmid DNA (1µg of a luciferase reporter vector based on the pGL2-Basic Vector, 0.5µg of an expression vector and 0.5µg of a Renilla luciferase plasmid as an internal control) and 3µl of FuGENE® 6 reagent in 0.1 ml of DMEM per dish. Reporter assays were performed using a dual luciferase assay system. (4279)

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FASEB J. 14, 2486-94. High-efficiency transient transfection of endothelial cells for functional analysis. 2000

Kovala, A.T., Harvey, K.A., McGlynn, P,. Boguslawski, G., Garcia, J.G., and English, D.

Notes: This study describes development of a method for transfection of endothelial cells. Bovine pulmonary aortic endothelial cells (BPAECs) were used to demonstrate the usefulness of the technique. Cells were cotransfected with a plasmid encoding GFP and a plasmid containing the gene of interest. Cells were first sorted by FACS analysis based on GFP expression and then analysed for the presence of the gene of interest. FuGENE® 6 was used for transfection. Cells were cultured in 100mm dishes and 5-10µg of DNA was used for transfection. (4307)

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J. Biol. Chem. 275, 9841-8. Inhibition of tumor necrosis factor-alpha transcription in macrophages exposed to febrile range temperature. A possible role for heat shock factor-1 as a negative transcriptional regulator. 2000

Singh, I.S., Viscardi, R.M., Kalvakolanu, I., Calderwood, S., and Hasday, J.D.

Notes: These authors used the FuGENE® 6 reagent to transiently transfect RAW 264.7 mouse macrophage cells with 4.5µg DNA at a 3.3:1 transfection reagent:DNA ratio. Cells were grown in 60mm dishes prior to transfection. (4394)

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Nat. Cell Biol. 2, 879-887. Molecular basis of transdifferentiation of pancreas to liver. 2000

Shen, C.N., Slack, J.M., and Tosh, D.

Notes: This paper showed that pancreatic cells can be converted into hepatocytes by treatment with dexamethasone. The conversion occurred in a pancreatic cell line, AR42J-B13 and in mouse organ cultures. FuGENE® 6 was used to transiently and stably transfect the rat pancreatic cancer cell line. 1.5 µg DNA was used for stable transfection, and 2µg was used for transient transfection. (4360)

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Proc. Natl. Acad. Sci. USA 97, 13275-80. Prostacyclin-mediated activation of peroxisome proliferator-activated receptor delta in colorectal cancer. 2000

Gupta, R.A., Tan, J., Krause, W.F., Geraci, M.W., Willson, T.M., Dey, S.K., and DuBois, R.N.

Notes: In this paper, FuGENE® 6 reagent was used to transiently transfect U2OS cells with 0.45µg DNA at a 3:1 FuGENE:DNA ratio. Cells were plated at 5 x 10e5 cells/well in 24-well plates. (4399)

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J. Virol. 74, 5776–87. The long terminal repeat of Jaagsiekte sheep retrovirus is preferentially active in differentiated epithelial cells of the lungs. 2000

Palmarini, M., Datta, S., Omid, R., Murgia, C. and Fan. H.

Notes: In general, transient transfections were performed using 2 × 105 to 4 × 105 cells plated in 6-well plates 24 hours before transfection, with a total of 2µg of plasmid (1µg of reporter vector and 1µg of control) using FuGENE® 6 reagent. The reporter vectors were based on the pGL3-Basic and pGL3-Promoter Vectors.

For MLE-15 (mouse lung), mtCC1-2 (mouse Clara cell), NIH-3T3 (mouse embryo), TCMK (mouse kidney), ST3, (mouse thymus) CP-MRI (sheep choroid plexus) and CP-ATCC (sheep choroid plexus) transfections used 0.5µg of reporter plasmid and 0.5µg of pRL-TK or 50ng pRL-null Vector. Reporter plasmid activity was assessed using the Dual-Luciferase® Assay System. (4271)

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J. Biol. Chem. 275, 38589–38596. Identification of a functional nuclear export sequence in BRCA1. 2000

Rodríguez, J.A. and Henderson, B.R. 

Notes: Several cancer-associated proteins, such as the tumor suppressor p53 and the oncoproteins c-ABL and hdm2, have been shown to shuttle between the nucleus and the cytoplasm. Due to this information and the controversy regarding BRCA1 localization, the authors wanted to determine whether BRCA1 is also capable of nuclear-cytoplasmic shuttling.

To this end, a plasmid encoding untagged BRCA1 was created by subcloning the full-length BRCA1 cDNA into a vector as a NotI/ClaI fragment. To generate pYFP-BRCA1, a DNA fragment encoding the yellow fluorescent protein (YFP) was amplified by polymerase chain reaction (PCR) and inserted in frame at the end of the BRCA1 cDNA in the plasmid, using the unique NotI restriction site.

Human breast cancer cell lines T47D and MCF-7, the human breast epithelial cell line HBL-100, and mouse NIH3T3 fibroblasts were seeded onto glass coverslips and transfected at 50–70% confluency with 0.5–2µg of plasmid DNA using the FuGENE® 6 Transfection Reagent. After 48 hours, cells were fixed and processed for fluorescence microscopy. (4283)

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J. Biol. Chem. 275, 26277–26284. Presenilin 1 regulates pharmacologically distinct gamma -secretase activities. Implications for the role of presenilin in gamma -secretase cleavage. 2000

Murphy M.P., Uljon S.N., Fraser P.E., Fauq A., Lookingbill H.A., Findlay K.A., Smith T.E., Lewis P.A., McLendon D.C., Wang R. and Golde T.E.

Notes: To generate stably-transfected lines, six-well plates of either Chinese hamster ovary (CHO), human embryonic kidney 293 (HEK), or human neuroglioma (H4) cells were transfected with 1µg of plasmid DNA preincubated with 3µl of FuGENE® 6 Transfection Reagent in serum-free medium overnight. Media were then replaced with media supplemented with 10% fetal bovine serum and 800mg/ml Hygromycin B. Transient transfections were performed in an identical manner, albeit without hygromycin selection. (4282)

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J. Biol. Chem. 274, 30778–30783. DECAY, a novel Drosophila caspase related to mammalian caspase-3 and caspase-7. 1999

Dorstyn, L., Read, S.H., Quinn, L.M., Richardson, H. and Kumar, S.

Notes: NIH3T3 cells were grown in Dulbecco's modified Eagle's medium with 10% fetal calf serum and 293T cells in RPMI with 10% fetal calf serum. For cell death assays, 2 × 105 cells were plated in each 35mm dish the day before transfection. pcDNA3-decay, pcDNA3-decay (C150G) or empty vector (1.5μg) were cotransfected with 0.5μg of a β-galactosidase expression plasmid (pEF-βgal). All transfections were carried out using FuGENE® 6 Transfection Reagent according to manufacturer's instructions. (4304)

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J. Cell Biol. 147, 77–88. ErbB4 signaling in the mammary gland is required for lobuloalveolar development and Stat5 activation during lactation. 1999

Jones, F.E., Welte, T., Fu, X.Y. and Stern, D.F.

Notes: 293T cells were transfected using FuGENE® 6 Transfection Reagent according to the manufacturer's instructions. Cells were grown to 25% confluency in 100mm tissue culture dishes, then transfected with 500μl of growth medium without serum, containing 10μl of FuGENE® 6 reagent and 2μg of each plasmid, for a total of 4μg. The cells were incubated with transfection mixture for 48 hours in a humidified incubator at 37°C with 5% CO2. (4305)

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J. Exp. Med. 190, 1679–1688. FLICE-inhibitory protein expression during macrophage differentiation confers resistance to fas-mediated apoptosis. 1999

Perlman, H., Pagliari, L.J., Georganas, C., Mano, T., Walsh, K. and Pope, R.M.

Notes: U937 human leukemic monocyte lymphoma cells were cultured at a density of 3 × 106 in 100mm plates for transient transfection. The cells were transfected with 10µg of plasmid DNA using FuGENE® 6 Transfection Reagent at a 1:5 ratio of DNA to reagent. After transfection, the cells were washed, incubated in medium for 12 hours before a 12-hour treatment with antibody and analysis by flow cytometry. (4405)

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J. Biol. Chem. 274, 32091–32098. The nuclear factor-kappaB engages CBP/p300 and histone acetyltransferase activity for transcriptional activation of the interleukin-6 gene promoter. 1999

Vanden Berghe, W., De Bosscher, K., Boone, E., Plaisance, S. and Haegeman, G.

Notes: In this paper, the authors continued their study of transcriptional mechanisms of TNF- and STS-mediated IL-6 gene activation, focusing on the functional interaction between TNF- or STS-responsive DNA-bound factors and the cofactor CBP/p300 in the IL-6 promoter context. The relation between cofactor-dependent HAT activity and IL-6 promoter stimulation was further explored with a HAT-defective p300 variant and with the potent HDAC inhibitor trichostatin A (TSA). They then extended their observations to additional NF-κB-containing promoters.

Transient transfection of HEK293T cells was performed using FuGENE® 6 Transfection Reagent according to the manufacturer's instructions. Approximately 5 × 104 HEK293T cells were seeded in 24-well plates 24 hours before transfection. After mixing FuGENE® 6 reagent with the DNA plasmids of interest, transfection mixtures (containing a total amount of 0.5µg of DNA) were added to each well and incubated with the cells for 24 hours, after which medium was refreshed and cells were further used in induction experiments. (4303)

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J. Biol. Chem. 273, 25292-300. A novel distal enhancer confers chorionic expression on the human renin gene. 1998

Germain, S., Bonnet, F., Philippe, J., Fuchs, S., Corvol, P., and Pinet, F.

Notes: In this study, several cell types were transfected with DNA using FuGENE® 6 reagent. Cells included COS-7 cells, primary cultures of chorionic cells and Calu-6 cells (derived from a pulmonary carcinoma), and As4.1 cells (ATCC CRL2193)—a mouse renin-expressing cell line. All cells were plated in 12-well plates. For chorionic cells, 3.4μl of FuGENE® reagent, 1.5μg reporter construct and 200ng of control DNA were used. For Calu-6 and COS-7 cells, 200ng of reporter construct and 30ng of control DNA were used with 0.92μl (Calu-6) or 0.46μl (COS-7) of FuGENE® reagent. For As4.1 cells, 4.6μl of FuGENE® reagent, 2μg of reporter construct and 300ng of control DNA were used. (4323)

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