RNA-seq of H2afj-ko and isogenic WT C57/Bl6-N mouse embryonic fibroblasts in proliferation and after etoposide-induced senescence.
Ontology highlight
ABSTRACT: Determine the effect of knocking out the H2afj gene on the transcriptome of MEFs induced into senescence with etoposide. MEFs were produced from individual H2A.J-ko or isogenic WT C57BL/6-N embryos. 3 different female WT and H2A.J-ko MEFs were used for the transcriptome analyses. MEFs were cultivated in DMEM + Gluta-Max (Gibco 31966) + 10%FBS + pen/strep in a 5% carbon dioxyde and 5% oxygen incubator. MEFs were passed twice before inducing senescence by treatment with 2.5 µM etoposide for 6 days (with media changed after 3 days), followed by 3 days incubation in fresh medium without etoposide.
Project description:The intent of the experiment was to test the effect of overexpressing H2A.J-V11A, H2AJ-S123E, H2A.J-S123A, and H2A.J-CterH2A mutants, as well as WT-H2A.J and canonical H2A-type1, on the transcriptome of WI-38hTERT fibroblasts. pTRIPz based lentiviral constructs placing these coding sequences under tetON control were used to produce stable WI38hTERT cell lines for each construct. Expression of the constructs in proliferating cells was induced with 100 ng/ml doxycycline for è days. Transcriptomes were also compared with control proliferating WI-38hTERT and WI-38hTERT cells induced into senescence by incubation with 20 µM etoposide for 2 weeks followed by 1 week without etoposide.
Project description:The experiment is intended to test the effect of the Cripsr/Cas9 deletion of the H2AFJ gene encoding the histone variant H2A.J on the transcriptome of luminal breast cancer T47D cells in conditions of proliferation, and after proliferative arrest induced by treating the cells with 5 µm tamoxifen for 4 days. Two independent H2A.J-KO cell lines were analyzed: KO1 and KO18.
Project description:Study the role of H2A.J in gene expression in senescent human fibroblasts by comparing RNA-seq of WI-38hTERT fibroblasts expressing either an sh2-H2AFJ or sh-NoTarget RNAs. Senescence was induced by treating cells with 20 M etoposide for 2 weeks followed by one further week of incubation without etoposide. 3 biological replicates of sh2-H2AFJ and sh-NoTarget were sequenced.
Project description:The senescence of mammalian cells is characterized by a proliferative arrest in response to stress and the expression of an inflammatory phenotype. We discovered that H2A.J, a poorly studied H2A variant found only in mammals, accumulates in human fibroblasts in senescence with persistent DNA damage. Knock-down of H2A.J interfered with the derepression of a set of inflammatory genes that contribute to the senescent-associated secretory phenotype (SASP), and over-expression of H2A.J increased the expression of some of these genes in proliferating cells. H2A.J accumulation may thus promote the signaling of senescent cells to the immune system. 41 samples analysed
Project description:Primary mouse embryonic fibroblasts (MEFs) from wt and MK2/3 KO mice were treated with 20µM etoposide. Primary MEFs from wt and MK2/3 KO mice were treated with 20µM etoposide and RNA extracted after 1h and 6h.
Project description:The intent of the experiment was to identify differentially expressed RNAs comparing WI-38hTERT/sh3-H2AFJ and WI-38hTERT/sh-NT fibroblasts in proliferation and etoposide-induced senescence.
Project description:The intent of the experiment was to identify differentially expressed micro-RNAs comparing WI-38hTERT/sh3-H2AFJ and WI-38hTERT/sh-NT fibroblasts in proliferation and etoposide-induced senescence.
Project description:The intent of the experiment was to compare H3-K4me3 peaks on the chromatin of WI-38hTERT/sh3-H2AFJ and WI-38hTERT/sh-NT fibroblasts in proliferation and etoposide-induced senescence.
Project description:The intent of the experiment was to compare RNA polymerase II occupancy on the chromatin of WI-38hTERT/sh3-H2AFJ and WI-38hTERT/sh-NT fibroblasts in proliferation and etoposide-induced senescence.
Project description:The senescence of mammalian cells is characterized by a proliferative arrest in response to stress and the expression of an inflammatory phenotype. We discovered that H2A.J, a poorly studied H2A variant found only in mammals, accumulates in human fibroblasts in senescence with persistent DNA damage. Knock-down of H2A.J interfered with the derepression of a set of inflammatory genes that contribute to the senescent-associated secretory phenotype (SASP), and over-expression of H2A.J increased the expression of some of these genes in proliferating cells. H2A.J accumulation may thus promote the signaling of senescent cells to the immune system.