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. Author manuscript; available in PMC: 2020 Jul 30.
Published in final edited form as: Cell Rep. 2020 Jul 14;32(2):107884. doi: 10.1016/j.celrep.2020.107884

Figure 7. Inhibition of CDK7 by THZ1 Sensitizes Cancer Cells to PARPi-Induced DNA Damage and Cell Death.

Figure 7.

(A) DNA damage in MDA-MB-231, OVCAR5, and DU145 cells treated with DMSO, THZ1, olaparib, or THZ1 combined with olaparib, measured by the comet assay. Scale bars, 10 μm.

(B) Extent of DNA damage, quantified by the tail moment in the comet assay. Data represented as means ± SD, n = 100 cells measured, *p < 0.05 determined by two-tailed Student’s t test.

(C) Representative images of γH2AX foci in MDA-MB-231, OVCAR5, and DU145 cells treated with DMSO, THZ1, olaparib, or THZ1 combined with olaparib. Scale bars, 5 μm. DAPI, 4′,6-diamidino-2-phenylindole.

(D) Quantification of the number of γH2AX-foci-positive cells in MDA-MB-231, OVCAR5, and DU145 cells treated with DMSO, THZ1, olaparib, or THZ1 combined with olaparib. Data represented as means ± SD, n = 3 biological replicates, *p < 0.05 determined by two-tailed Student’s t test.

(E) Western blot analysis of γH2AX in MDA-MB-231, OVCAR5, and DU145 cells treated with DMSO, THZ1, olaparib, or THZ1 combined with olaparib.

(F) Percentage of annexin-V-positive cells in MDA-MB-231, OVCAR5, and DU145 cells treated with DMSO, THZ1, olaparib, or THZ1 combined with olaparib. Data represented as means ± SD, n = 3 biological replicates, *p < 0.05 determined by two-tailed Student’s t test.

(G) Caspase-3/7 activity, in MDA-MB-231, OVCAR5, and DU145 cells treated with DMSO, THZ1, olaparib, or THZ1 combined with olaparib, was measured by a caspase-3/7 assay. Data represented as means ± SD, n = 3 biological replicates, *p < 0.05 determined by two-tailed Student’s t test.

(H) Cleaved-PARP1, cleaved-caspase 9, and cleaved-caspase 3 expression assessed by western blots in MDA-MB-231, OVCAR5, and DU145 cells treated with DMSO, THZ1, olaparib, or THZ1 combined with olaparib are shown. Actin served as a loading control.

(I) Percentage of annexin-V-positive MDA-MB-231, OVCAR5, and DU145 cells treated with DMSO or olaparib combined with CDK7 siRNAs. Data represented as means ± SD, n = 3 biological replicates, *p < 0.05 determined by two-tailed Student’s t test.