Project description:To better understand the mechanism by which MAGEA3 contributes to HCC progression, we conducted RNA sequencing in the PLC5 HCC cell line after 72 hours of treatment with scramble or sh8375 in triplicate. RNA-sequencing was conducted on poly-A enriched RNA, 100 bp single reads using an Illumina HiSeq2500 instrument. Libraries were constructed using the TruSeq RNA Library Prep Kit v2. Raw sequencing reads were mapped to the GRCh38 reference genome (USCS) using STAR (2.4.2g1). Aligned reads were mapped to GRCh38 genetic features using featureCounts from the subRead package with default settings. Data analysis included differential gene expression between conditions and gene set enrichment analysis.
Project description:The cumulus cells niche that surrounds the oocyte is essential for its maturation and presumably for the oocyte to acquire its competence to confer pluripotency. The cells cultured from the human oocyte cumulus niche (hCC) could be used as feeders for the propagation of human pluripotent stem cells in vitro. We used microarrays to establish the molecular portrait of the hCC at different passages
Project description:Purpose: Next-generation sequencing (NGS) has revolutionized systems-based analysis of cellular pathways. The goals of this study are to compare NGS-derived H2O2-treated HCC transcriptome profiling (RNA-seq) to NGS-derived mock-treated HCC transcriptome profiling. Methods: HCC mRNA profiles of mock-treated Huh7 cells and H2O2-treated Huh7 cells were generated by deep sequencing, in duplicate, using Illumina Pipeline (CASAVA) v1.8.2. The sequence reads that passed quality filters were analyzed at the transcript isoform level with TopHat followed by Cufflinks. qRT–PCR validation was performed using TaqMan and SYBR Green assays Results: RNA-seq data validated with qRT–PCR. RNA-seq data had a linear relationship with qRT–PCR for more than four orders of magnitude and a goodness of fit (R2) of 0.90. Conclusions: We conclude that our study represents the first detailed analysis of H2O2-treated HCC cells transcriptomes compared to mocke-treated HCC cells, with biologic replicates, generated by RNA-seq technology.
