Fig. 6

Disruption of EHMT1/2 in PEO1-OR cells impairs DNA repair and significantly alters gene expression. a Immunofluorescence staining for DNA damage marker γH2AX was performed on PEO1-OR shControl or shEHMT1/2 double knockdown cells. Nuclei were counterstained with DAPI. b Staining for γH2AX as in (a), except cells are PEO1-OR treated for 72 h with 1 μM UNC0638, UNC0642, or DMSO control. For (a, b), the percentage of γH2AX positive nuclei are plotted (mean ± SD, ≥ 200 nuclei per test, n = 3 slides per condition, unpaired t test). c, d Scramble shRNA and EHMT1/2 double knockdown PEO1-OR cells were assayed for HR and NHEJ capacity using a two-plasmid GFP reporter assay. GFP+ cells percentage was determined by flow cytometry. e, f PEO1-OR cells were treated for 72 h with 1 μM UNC0638 or UNC0642 DMSO control, then assayed for HR and NHEJ capacity as in (c, d). GFP+ cell percentage is plotted (mean ± SD, 50,000 cells analyzed per test, n = 3, unpaired t test). g PEO1-OR cells were treated for 72 h with UNC0642, then the transcriptome was analyzed by RNA-Seq and compared to the transcriptome of four untreated PEO1-OR clones. Genes are plotted by fold change and p value. Red dots represent genes with false discovery rate (FDR) < 0.001 and absolute value of Log2 fold change > 0.5