Project description:in vivo restoration of aged bone marrow mononuclear cells by microRNA: effect on cellular aging Treatment groups were (1) no microRNA, (2) microRNA-619 (miR-619), (3) a combination of microRNAs-619, -1303, 4497 (miR-combo), or (4) a control microRNA expected to have no effect (control RNA)
Project description:Three HCT116 cell lines (HCT WT, HCT p21-/-, HCT p53-/-) were analyzed by RT-qPCR array analysis for chromatin modification enzymes to identify potential target genes, that depend on the cell cycle regulator p21 and that are indpendent of p53. HCT116 cell lines (HCT WT, HCT p21-/-, HCT p53-/-) were cultured at 37°C in a humidified atmosphere with 5% CO2. Cells were harvested at a confluency of ~60-80% and RNA was isolatd using QIAzol an Rneasy Kits from Qiagen. cDNA was produced from equal amounts of RNA using the RT² First Strand Ki from Qiagen and RT-qPCR analysis was performed following the manufacturer's instructions using the Epigenetic Chromatin Modification Enzymes RT² Profiler PCR Array from Qiagen. The CFX96TM Real-Time System and the C1000TM Thermal Cycler from Bio-Rad were used to perform the RT-PCR runs. The three different cell lines were analyzed in three biological triplicates. The mean expression Ct values of the target genes were normalized against the mean Ct value for GAPDH to calculate the Fold Change values. The Raw Data was processed using the RT² Profiler PCR Array Data Analysis version 3.5 software from SABiosciences.
Project description:Two HCT116 cell lines (HCT WT, HCT p21-/-) were analyzed by RT-qPCR array analysis for genes associated with Epithelial to Mesenchymal Transition (EMT) to identify potential target genes, that depend on the cell cycle regulator p21. HCT116 cell lines (HCT WT, HCT p21-/-) were cultured at 37°C in a humidified atmosphere with 5% CO2. Cells were harvested at a confluency of ~60-80% and RNA was isolatd using QIAzol an Rneasy Kits from Qiagen. cDNA was produced from equal amounts of RNA using the RT² First Strand Ki from Qiagen and RT-qPCR analysis was performed following the manufacturer's instructions using the Epithelial to Mesenchymal Transition RT² Profiler PCR Array from Qiagen. The CFX96TM Real-Time System and the C1000TM Thermal Cycler from Bio-Rad were used to perform the RT-PCR runs. The two different cell lines were analyzed in three biological triplicates. The mean expression Ct values of the target genes were normalized against the mean Ct value for B2M to calculate the Fold Change values. The Raw Data was processed using the RT² Profiler PCR Array Data Analysis version 3.5 software from SABiosciences.
Project description:In vivo restoration of aged bone marrow mononuclear cells by micrRNA: effect on cellular senesence Treatment groups were (1) no microRNA, (2) microRNA-619 (miR-619), (3) a combination of microRNAs-619, -1303, 4497 (miR-combo), or (4) a control microRNA expected to have no effect (control RNA)
Project description:mRNA levels of the known 91 FoxO1 target genes were evaluated with RT2 Profiler PCR array in cardiomyocytes transduced with LacZ, Mst1, FoxO1, FoxO1+Mst1, or FoxO1+DN-Mst1 genes were evaluated with RT2 Profiler PCR arrays.
