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. 2020 Jul 23;21(15):5239. doi: 10.3390/ijms21155239

Figure 7.

Figure 7

miR-711 inhibition attenuates IR-induced DNA damage markers, p53 activation, and neuronal apoptosis and senescence markers. Western blot for Ph-ATM(Ser1981), γ-H2A.X and H2A.X (A). (B) Ph-ATM(Ser1981) (F(10,22) = 173, p < 0.0001 at 30 min, 1 h, 3 h and 6 h after IR with miR-ve inhibitor, compared to control for miR-711 inhibitor, compared to miR-ve inhibitor, p < 0.0001at 1 h and 3 h, p = 0.0001 at 6 h), γH2A.X normalized to β-actin (F(10,22) = 281.7, p < 0.0001 for all time points except 24 h after IR with miR-ve inhibitor, compared to non-irradiated control; for miR-711 inhibitor, compared to miR-ve inhibitor, p < 0.0001 for 1 h and 3 h, p = 0.0005 for 6 h), γH2A.X normalized to H2A.X (F(10,22) = 73.18, p < 0.0001 for all time points except 24 h after IR with miR-ve inhibitor, compared to control for miR-711 inhibitor, compared to miR-ve inhibitor, p = 0.0017 at 1 h, p < 0.0001 at 3 h and 6 h). H2A.X (F(10,22) = 138.8, p < 0.0001 for 1 h and 3 h after IR with miR-ve inhibitor, compared to control; for miR-711 inhibitor, compared to miR-ve inhibitor, p < 0.0001 for 1 h and 3 h after IR with miR-ve inhibitor, compared to control; for miR-711 inhibitor, compared to miR-ve inhibitor, p < 0.0001 at 30 min, p = 0.0029 at 1 h). Western blots for Ph-ATR(Ser428), Ph-p53(Ser15), p53, p21 and β-actin (C). (D) Ph-ATR(Ser428)(F(8,18) = 108.4, p < 0.0001 at 30 min, 3 h and 6 h, p = 0.0017 at 24 h after IR with miR-ve inhibitor, compared to control; for miR-711 inhibitor, compared to miR-ve inhibitor, p < 0.0001 at 3 h and 6 h), Ph-p53(Ser15) normalized to β-actin (F(8,18) = 302, p < 0.0001 at 3 h and 6 h after IR with miR-ve inhibitor, compared to control; for miR-711 inhibitor, compared to miR-ve inhibitor, p < 0.0001 at 3 h and 6 h), Ph-p53(Ser15) normalized to p53(F(8,18) = 392.6, p < 0.0001 at 3 h and 6 h after IR with miR-ve inhibitor, compared to control; for miR-711 inhibitor, compared to miR-ve inhibitor, p < 0.0001 at 3 h and 6 h), p21(F(8,18) = 45.58, p < 0.0001 at 3 h and 6 h, p = 0.0410 at 24 h after IR with miR-ve inhibitor, compared to control; for miR-711 inhibitor, compared to miR-ve inhibitor, p = 0.0001 at 6 h). n = 3/group. qPCR quantification of p21 and p53 mRNAs (E). p21 mRNA (F(10,25) = 111, p = 0.0342 at 30 min, p < 0.0001 at 3 h and 6 h after IR with miR-ve inhibitor, compared to control; for miR-711 inhibitor, compared to miR-ve inhibitor, p < 0.0001 at 3 h and 6 h), p53 mRNA (F(10,25) = 5.062, no significant changes). n = 3/group. * p < 0.05, **** p < 0.0001 vs. control, ^^^ p < 0.001, ^^^^ p < 0.0001 vs. corresponding 8Gy + miR-ve inhibitor group. miR-711 inhibition attenuates irradiation-induced downregulation of DNA repair molecules Rad50 and Rad54l2.