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. 2020 Feb 10;10(7):3351–3365. doi: 10.7150/thno.41219

Figure 2.

Figure 2

Olaparib increases DNA damage induced by TMZ by inhibiting the ATRX mediated PARP1 stabilization. (A) GSEA of the apoptotic signaling pathway was performed in ATRX NC and ATRX KO cells. (B) Differentially expressed genes of the apoptotic signaling pathway between ATRX NC and ATRX KO cells and their interaction with PARP1. (C) Western blotting of PARP1, c-PARP1 and γ-H2AX of ATRX NC and ATRX KO cells treated with 200 μM TMZ and/or 1 μM olaparib. TMZ and olaparib were added to culture medium for 72 hours. (D) CASP3/7 activity of ATRX NC and ATRX KO cells treated with TMZ and/or olaparib. TMZ and olaparib were added to culture medium for 72 hours. (E) TMZ and olaparib were added to culture medium at final concentrations of 200 μM and 1 μM, respectively, for 72 hours. γ-H2AX is shown in red and nucleus in blue. Bar plots showed the statistics of immunofluorescence assays in HG7R. Scales: 20 μm. (F) Comet assays measuring the DNA damage status in ATRX NC and ATRX KO cells treated with TMZ or combination of TMZ and olaparib in HG7R. Scales: 100 μm. Error bars indicate mean ± SD. *** P < 0.001, Student's t-test.