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Fig. 4. | Clinical Epigenetics

Fig. 4.

From: Histone methyltransferases EHMT1 and EHMT2 (GLP/G9A) maintain PARP inhibitor resistance in high-grade serous ovarian carcinoma

Fig. 4.

Knockdown or inhibition of EHMT1/2 resensitizes PEO1-OR cells to olaparib. a PEO1-OR cells were transduced with lentivirus carrying shRNAs targeting both EHMT1 and EHMT2 or Scramble control. Knockdown of EHMT1 and EHMT2 mRNA was analyzed by RT-qPCR (mean ± SD, n = 3, unpaired t test). b Knockdown of EHMT1 and EHMT2 protein was analyzed by immunoblot. Actin was used as loading control. c Histone extracts of double knockdown cells were analyzed by immunoblot for H3K9me2 and compared to Scramble control cells. Total H3 was used as loading control and for densitometry analysis. d Resensitization of EHMT1/2 double knockdown PEO1-OR cells was analyzed by dose response colony formation assays and compared to Scramble control cells. Dose response curves are graphed with IC50 indicated. e Cells from dose response assay were stained with crystal violet. Colony formation is plotted (mean ± SD of three wells, unpaired t test). f PEO1-OR cells were treated for 72 h with DMSO control or EHMT1/2-specific methyltransferase inhibitor UNC0642 at two-fold dilutions from 2 μM to 125 nM. H3K9me2 was analyzed by immunoblot or immunofluorescence. To confirm drug specificity, H3K27me3 was analyzed my immunoblot. Total H3 was used as loading control. IF images are representative from DMSO control and 500 nM treatment. g Resensitization of PEO1-OR cells co-treated with 1 μM UNC0642 was analyzed by dose response colony formation assays and compared to DMSO control treatment. Colony formation is plotted (mean ± SD of four wells, unpaired t test)

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