Project description:Microarray data from G2-synchronized p53(+) and p53(-) fibroblasts before and after 3 h release from cell cycle blockade in the presence of 5 µM sodium arsenite. Keywords: Gene induction
Project description:Microarray data from G2-synchronized p53(+) and p53(-) fibroblasts before and after 3 h release from cell cycle blockade in the presence of 5 uM sodium arsenite. Experiment Overall Design: Cells expressing p53 from a tet-off regulated construct were synchronized in G2 with a two-step procedure using 24 h aphidicolin treatment for initial G1 synchronization and 12 h of Hoechst 33342 to effect a G2 blockade. During Hoechst treatment, tetracycline was added to suppress p53 in half the cultures. Cells were then released into media containing 5 µM sodium arsenite and the appropriate concentration of tetracycline to maintaining p53 expression. mRNAs were collected at 0 h and 3 h after release from G2 synchrony.
Project description:For senescence induction, HCA2 humn skin fibroblasts were synchronized at G2 phase by treatment with 9 uM of RO3306 (Roche) for 24 hrs, treated with 9 uM of RO3306 and 5 uM of Nutlin3a (Sigma-Aldrich) for 8 hrs, and then with 5 uM of nutlin3a for 18 hrs. Cells were then treated with 100 nM of BI-2536 for 9 days to eliminate proliferating cells, and cultured in normal media for 9 days. The senescent and non-senescent (young) cells were collected, washed with PBS, snap-frozen, and analyzed with a iMPAQT method.
Project description:Arsenic is a potent environmental toxin and a cause of numerous health problems. Most studies have assumed that arsenic-induced changes in mRNA levels result from effects on gene transcription. The influence of arsenic on post-transcriptional regulation, another important locus of gene expression control, has remained largely unexplored. To evaluate the prevalence of changes in mRNA stability in response to arsenic in human fibroblasts, we used microarray analyses to determine changes in steady state mRNA levels, and their decay rates, following 24 hour exposure to non-cytotoxic concentrations of sodium arsenite (1 µM). We conclude that arsenite modification of mRNA stability is relatively uncommon, but in some instances can result in significant changes in gene expression. Human BJ diploid foreskin fibroblasts were used in the study. The decay rates of transcripts were determined using actinomycin D to stop transcription after sodium arsenite or water treatment for 24 h. Actinomycin D was added into the culture medium at a final concentration of 5 µg/ml, and treated cells were then harvested at 0, 1, 2, 3 and 4 h for RNA extraction.
Project description:Arsenic is a potent environmental toxin and a cause of numerous health problems. Most studies have assumed that arsenic-induced changes in mRNA levels result from effects on gene transcription. The influence of arsenic on post-transcriptional regulation, another important locus of gene expression control, has remained largely unexplored. To evaluate the prevalence of changes in mRNA stability in response to arsenic in human fibroblasts, we used microarray analyses to determine changes in steady state mRNA levels, and their decay rates, following 24 hour exposure to non-cytotoxic concentrations of sodium arsenite (1 µM). We conclude that arsenite modification of mRNA stability is relatively uncommon, but in some instances can result in significant changes in gene expression.
Project description:We performed RNA-seq in cells synchronized to S or G2 phase to identify genes that were transcriptionally regulated by ATR activity.
