Project description:We used siRNA to individually knock down the human phospholipid transfer proteins STARD2, STARD7 and STARD10 in HepG2 cells and measured the impact on the cellular phospholipid metabolism. We used DIA proteomics to quantify the selective reduction in protein expression levels.
Project description:HNF4a is an important liver transcription factor that regulates at least a thousand genes in the liver. Here we used expression profiling in HepG2 cells, a hepatocellular carcinoma cell line, in which HNF4a was knocked down by RNAi to identify some of those target genes. This dataset accompanies the article in Hepatology 2010 Feb;51(2):642-53. Integrated approach for the identification of human hepatocyte nuclear factor 4alpha target genes using protein binding microarrays by Bolotin E, Liao H, Ta TC, Yang C, Hwang-Verslues W, Evans JR, Jiang T, Sladek FM. RNA interference (RNAi) against HNF4a2 was performed in HepG2 cells using small, interfering RNAs (siRNAs) corresponding to nucleotides +179 to +197 of human HNF4A (NM_178849, sense siRNA: 5'-UGUGCAGGUGUUGACGAUGdTdT-3', antisense siRNA 5'-CAUCGUCAACACCUGCACAdTdT-3') (Dharmacon, Lafayette, CO). Total RNA was extracted with Trizol (Life Technologies, Carlsbad, CA) and reverse transcribed with the Reverse Transcription System (Promega, Madison, WI). Polymerase chain reaction (PCR) amplification was performed in the linear range (see Supporting Table 3B for a list of PCR primers). Expression profiling analysis was performed with Affymetrix oligonucleotide arrays (HGU133 Plus 2.0) using RNA from control (PGL3 siRNA) or treated (HNF4a siRNA) HepG2 cells
Project description:To determine the effects of UBL5 on genome-wide gene expression, we used UBL5 siRNA to knock down UBL5 in HepG2 and Huh7 cells. We then performed gene expression profiling analysis using data obtained from RNA-seq.
Project description:Six1, Six4 and Myogenin are transcription factors that are known to be required for skeletal myogenesis. Currently, very little is known about the genes targeted by Six1 and Six4. Gene expression profiling when one or both transcription factors were knock-down by siRNA was performed to identify genes affected by their absence. We also hypothesized that Six1 and Six4 can work in cooperation with the myogenic regulatory factor (MRFs) family of transcription factors, such as Myogenin. Therefore, we performed the same type of experiment where the myogenin was knocked-down by siRNA to identify genes that are possibly regulated by the Six1 or Six4 in conjunction with Myogenin.
Project description:Purpose: To build a differential transcriptome network in Smug1 knock-out HepG2 hepatocarcinoma cells. Methods: Transcriptome analysis by the RNA-seq via mRNA pull-down. Results: We constructed transcriptome from Smug1 knock-out cells by using RNA-seq analysis. Nucleosome and miRNA related genes were highly enriched in Smug1 KO HepG2 cells. Conclusions: Smug1 regulate the gene expression related with nucleosome assembly and histone function.
