Fig. 3: Effects of BKM120 and rucaparib as single agents or in combination on HR repair. | Cell Death & Disease

Fig. 3: Effects of BKM120 and rucaparib as single agents or in combination on HR repair.

From: BKM120 sensitizes glioblastoma to the PARP inhibitor rucaparib by suppressing homologous recombination repair

Fig. 3

A U251 and U87MG cells were transfected with DR-GFP plasmid and pCBASceI plasmid using Lipofectamine 3000. Cells were treated with BKM120 or/and rucaparib for 72 h and then collected and resuspended in ice-cold PBS. GFP intensity was analyzed by flow cytometry. B The percentage of GFP-positive cells represents HR repair efficiency. C, D U251 and U87MG cells were treated with BKM120 and/or rucaparib for 48 h. C Cells were collected for western blotting detection of DNA damage- and HR repair-related proteins, including BRCA1, BRCA2, RAD51, RRM2, p-ATR, p-CHK1, p-AKT, ATR, CHK1, and PAR. D Quantitative reverse transcription PCR analysis of BRCA1, BRCA2, and RAD51 expression in two cancer cell lines. Gene expression was normalized to 18 s rRNA. E, F Cells were treated with BKM120 or/and Rucaparib for 12/24 h. Representative images of immunofluorescence staining of RAD51 and γ-H2AX in GBM cells Cell nuclei were stained with DAPI. Scale bar: 20 μm. The percentage of cells with RAD51/γ-H2AX foci was shown in (F). All data are presented as mean ± SD (n = 3). *P < 0.05; **P < 0.01; ***P < 0.001.

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