Fig. 5
Disruption of EHMT1/2 in PEO1-OR cells alters cell cycle regulation, but not apoptosis or senescence. a Scramble shRNA and EHMT1/2 double knockdown PEO1-OR cells were treated with 3 μM olaparib for 72 h, then stained with Annexin V/PI followed by flow cytometry to assay for apoptosis and necrosis. b PEO1-OR cells were treated with 3 μM olaparib, 1 μM UNC0642, or a combination, and then assayed as in (a). For AnnexinV/PI analyses, cell % are plotted as violin plots with graphed means, n = 3. c β-galactosidase activity was assessed by detection of fluorescent C12FDG cleavage product in PEO1-OR cells treated as shown. Cells were analyzed using flow cytometry (mean ± SD, 50,000 cells per test, n = 3, unpaired t test). d PEO1-OR cells were treated as shown, then fixed and stained with propidium iodide for DNA content and analyzed by flow cytometry. Percentages of cells in G1, S, and G2 were calculated by the Dean-Jett-Fox method in FlowJo 10 (mean ± SD, 50,000 cells per test, n = 3). e Immunoblot analysis for cell cycle proteins using total protein lysates from PEO1-OR cells treated as shown