GSE129954
iMEF SDHC-loss stable line CUT&RUN
We have developed a tet-inducible SV-40 large T antigen-expressing lentivirus-immortalized mouse embryonic fibroblast (iMEF) cell culture model of mitochondrial electron transport chain (ETC) dysfunction involving complex II (succinate dehydrogenase; SDH) in which silencing gene rearrangement of the Sdhc floxed allele is driven by doxycycline-dependent expression of cre-recombinase from a tet-inducible promoter (R26M2rtTA/+;TetOcre;Sdhcfl/fl).  For comparison, we include an isogenic Sdhc wt control line (R26M2rtTA/+;TetOcre;Sdhcfl/wt) that retains one intact copy of Sdhc upon doxycycline exposure.  Following doxycycline induction of both cell lines, dilutional subcloning was employed to obtain stable Sdhc -/- (SDHC KO) and Sdhc +/- (Control) cell lines.  Sdhc gene rearrangement status for each clonally-derived cell line was confirmed by PCR.  Cell lines were grown in standard DMEM containing penicillin/streptomycin antibiotics (0.5 mg/mL), non-essential amino acids (100 micromolar each of glycine, alanine, asparagine, aspartic acid, glutamic acid, proline, and serine), sodium pyruvate (1 mM), and HEPES buffer (10 mM) at 21% O2 and 5% CO2.  Following cell line derivation and verification of Sdhc genetic status, chromatin epigenomic marks including H3K4me3, H3K27me2, H3K27ac, CTCF, acetyllysine, propionyllysine, and butyryllysine were characterized by CUT&RUN. Overall design: Included in this dataset are biological replicate CUT&RUN data for two SDHC KO (R26M2rtTA/+;TetOcre;Sdhc-/-) and  two control (R26M2rtTA/+;TetOcre;Sdhc+/-) cell lines derived by dilutional subcloning following induction of Sdhc gene rearrangement with doxycycline.  For each line, epigenomic marks including H3K4me3, H3K27me2, H3K27ac, CTCF, acetyllysine, propionyllysine, and butyryllysine were characterized by CUT&RUN.