Notes: Bafilomycin C1 has been shown to have cytotoxic effects specifically against cancer cells. Here, the mechanism of action of Bafilomycin C1 is investigated in human hepatocellular cancer SMMC7721 cells. Bafilomycin C1 shows mitochondrial membrane dysfunction, cell cycle arrest, and down-regulation of cell-cycle regulators. Additionally, bafilomycin C1 treatment lead to induction of apoptotic pathways. In preparation for real-time PCR, total RNA was extracted and cDNA was synthesized from the extracted total RNA using the GoScript™ Reverse Transcription System. Protease Inhibitor Cocktail was used to prepare cell lysates for Western blot analysis. (5086)

Notes: Researchers studied the liver stage of the Plasmodium parasite (which causes malaria), including RNA sequencing, parasite load and cell viability. (5035)

Notes: Reliaprep RNA cell miniprep system was used to extract RNA from Hematopoietic stem cells derived from pluripotent stem cells. Then qRT-PCR analysis of miRNA and reverse transcription was performed using non-Promega products and 1 μg of RNA was used in a 20 μl GoScript reverse transcription reaction for the analysis of gene expression by qRT-PCR. (4975)

Notes: RNA was extracted from the cells collected from differentiating hESC and hiPSC at days 0, 9, and 20 using the ReliaPrep RNA Cell Miniprep System. The RNA quality was evaluated using the Nanodrop 2000 spectrophotometer (Thermo Fisher Scientific, MA). One microgram of extracted RNA was converted into cDNA using reverse transcription (GoScript Transcription System). Quantitative real‐time polymerase chain reaction (qRT‐PCR) was carried out using the QuantStudio 7 Flex Real‐Time PCR System (Thermo Fisher Scientific, MA) and GoTaq qPCR Master Mix. pGL3 BRE Luciferase was used for plasmid lipofection to transfect the cells in each well of a 24-well plate. Cells that were transfected with empty vector (pGL3‐Basic) or BMP reporter (pGL3 BRE Luciferase) were cotransfected with empty Renilla vector (pRL‐Null). Luciferase activities were evaluated with a Dual‐Luciferase Assay System. (4982)

Notes: Primary pancreatic cancer cell lines were grown a microsphere by culturing in round bottom plates in the presence of methylcellulose. Spheroids were assessed for sensitivity to JQ1 using the CellTiter-Glo® 3D Cell Viability Assay. Cell lines expressing high levels of c-myc were sensitive to JQ1. RNA was isolated from cell lines and targeted expression analysis was performed by reverse transcription with the GoScript™ Reverse Transcription System followed by dye-based qPCR with the GoTaq® qPCR Master Mix on the AriaMx real-time PCR instrument (Agilent). (1825)

Notes: HEK 293 cells were treated with thapsigargin and expression of target mRNAs was examined. Total RNA was isolated with the ReliaPrep™ RNA Cell Miniprep System and reverse transcribed with the GoScript™ Reverse Transcription System followed by quantitation with the dye-based GoTaq® qPCR System. (4606)

Notes: The effect of SMYD3 and ERK2 on hepatitis B RNA expression was examine in transfected Huh-7.5 human hepatoma cells. Total RNA was isolated with the ReliaPrep™ RNA Cell Miniprep System and converted to cDNA with the GoScript™ Reverse Transcription System followed by dye-based qPCR analysis. The effect of SMYD3 and ERK2 on AP1-dependent and NF-κB-dependent gene expression in the cells was examined with the pGL4.44 [luc2P/AP1 RE/Hygro] and pGL4.32 [luc2P/NF-κB-RE/Hygro] Reporter vectors using the Dual-Luciferase® Reporter Assay System with a pRL-TK Renilla luciferase control. Reporter assays were read with a GloMax® Instrument. (4603)

Notes: In this study, researchers treated selected viral RNA samples with 1 unit of DNase I to deplete the presence of any genomic DNA in the extracts. The viral RNA genomic regions of interest were reverse transcribed into cDNA using oligo(dT) primer and AMV reverse transcriptase. (4887)

Notes: T47D and HB2 breast cancer cell lines were treated with doxorubicin and expressed in three target genes. Total RNA was isolated with the ReliaPrep™ RNA Cell Miniprep System, reverse transcribed with the GoScript™ Reverse Transcription System and analyzed with the dye-based GoTaq® qPCR System. (4612)

Notes: SR-786 NPM/ALK-positive human anaplastic large cell lymphoma cell line was treated with lobatin B, a plant-derived natural compound. Total RNA was isolated with the ReliaPrep™ RNA Cell Miniprep System and reversed transcribed into cDNA with the GoScript™ Reverse Transcription System. Levels of NPM/ALK and NF-κB expression was measured with the dye-based GoTaq® qPCR System. SR-786 cells were also analyzed for functioning metabolism with the CellTiter-Blue® Cell Viability Assay and apoptosis with the Apo-ONE™ Homogeneous Caspase-3/7 Assay. (4604)

Notes: Human Umbilical Vein Endothelial cells (HUVECs) were treated with IL-27, calprotectin, TNFα or combinations. Cell number per well was controlled by lysing the cells and measuring the LDH release with the CytoTox 96® Assay. Changes in gene expression were monitored by probe-based qPCR with cDNA made with the GoScript™ Reverse Transcription System using total RNA isolated using the ReliaPrep™ RNA Cell Miniprep System. Changes in protein levels upon treatment were evaluated by mass spec using Trypsin Gold for peptide analysis. (4598)

Notes: HEK 293 cells were transfected with various viral protein expression vectors and expression levels measured in the presence or absence of interferon-α using RT-qPCR. Total RNA was isolated from the cells using the ReliaPrep™ RNA Cell Miniprep System and converted to cDNA with the GoScript™ Reverse Transcription System prior to dye-based qPCR. (4613)

Notes: Total RNA was extracted from mucosa and denuded-detrusor layers of rat bladders with the ReliaPrep™ RNA Tissue Miniprep System and converted to cDNA using GoScript™ Reverse Transcription System prior to PCR amplification. (4608)

Notes: Rat renal DT and A3 cells were examined for differences in EGFR expression by RT-qPCR. Total RNA was isolated with the ReliaPrep™ RNA Cell Miniprep System and reverse transcribed with the GoScript™ Reverse Transcription System followed by dye-based qPCR analysis. (4617)

Notes: Primary splenocytes were isolated from mice immunized with an NS3 expression vector and examined for IFN-γ expression by RT-qPCR. Total RNA was isolated with the ReliaPrep™ RNA Cell Miniprep System and converted to cDNA with the GoScript™ Reverse Transcription System. The affect of wildtype and mutant NS3 on interferon-β promoter activity with an IFN-β promoter firefly luciferase vector and pRL-TK vector control was measured with the Dual-Luciferase® Reporter Assay System. LDH release was measured with the CytoTox 96® Cytotoxicity Assay in a cytotoxic T-lymphocyte assay using cultured splenocytes from the immunized mice. (4609)

Notes: Huh7.5 cells were infected with HCV and treated with the plant-derived chemicals. The expression of viral messages was analyzed by RT-qPCR. Total RNA was isolated with the ReliaPrep™ RNA Cell Miniprep System and reverse transcribed with the GoScript™ Reverse Transcription System before dye-based qPCR analysis. (4616)

Notes: Human Skin Progenitor (HSK) cells were cultured on hydrogels and either left alone or differentiated into various skin cell types. Gene expression changes were monitored by isolating total RNA from collagenase and hyaluronidase treated hydrogels with the ReliaPrep™ RNA Cell Miniprep System followed by cDNA synthesis with the GoScript™ Reverse Transcription System and qPCR with the dye-based GoTaq® qPCR System. (4602)

Notes: RNA was extracted from cultured A549 cells and used in RT-qPCR to analyze the effect of transfected siRNAs. (4444)

Notes: The authors examined cellular adaptation to increased salinity in Broussonetia papyrifera by measuring protein and mRNA levels of vacuolar H⁺-ATPase (V-H⁺-ATPase) subunits and the activities of V-H⁺-ATPase and vacuolar H⁺-pyrophosphatase. Relative expression levels of V-H⁺-ATPase subunits A, B, E and c in salt-stressed and control plants were determined by RT-PCR using actin as a normalization control gene. cDNA was synthesized using the GoScript™ Reverse Transcription System as described by the manufacturer’s protocol, then amplified by PCR for 25 cycles. The amplification products were analyzed and quantified by agarose gel electrophoresis and ethidium bromide staining. (4258)

Notes: The transcription factor Nanog is required for the maintenance of embryonic stem (ES) cell pluripotency. These authors showed that the Nanog N-terminal domain is regulated by post-transcriptional modification, and that alternative splicing generates Nanog variants with different capacities for maintaining an undifferentiated cell state. As part of their study, the authors used GoScript® Reverse Transcriptase to generate cDNA from RNA extracted from cell lines expressing different Nanog variants. The cDNA was used in RT-qPCR to quantify relative expression levels. (4184)