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

Figure 8.

Figure 8

miR-711 inhibition attenuates irradiation-induced downregulation of DNA repair molecules Rad50 and Rad54l2. qPCR quantification of RAD50 and RAD54l2 (A). Rad50 mRNA (F(10,22) = 43.49, p < 0.0001 at 30 min and 1 h, p = 0.0082 at 3 h after IR with miR-ve inhibitor, compared to control; for miR-711 inhibitor, compared to miR-ve inhibitor, p = 0.0039 at 3 h, p < 0.0001 at 6 h). Rad54l2 mRNA (F(10,22) = 39.1, p = 0.0005 at 30 min, p = 0.006 at 1 h, p = 0.0396 at 3 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, p = 0.0007 at 6 h). n = 3/group for all groups, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 vs. control; ^ p < 0.05, ^^ p < 0.01, ^^^ p < 0.001, ^^^^ p < 0.0001 vs. corresponding 8Gy + miR-ve inhibitor group. RCNs were exposed to 8Gy and, after 3 h, cells were collected and lysed. One part of the lysate from each sample was used for RNA isolation and qPCR analysis for levels of miR-711, RAD50 and RAD54l2 (B). miR-711 (T(4) = 3.856, p = 0.0091), Rad50 mRNA (T(4) = 7.944, p = 0.0007), Rad54l2 mRNA (T(4) = 0.74, p = 0.0002). The other part was subjected to RIP using Ago2 antibodies, followed by qPCR analysis for levels of miR-711, RAD50 and RAD54l2 in the RISC (C). miR-711 (T(4) = 9.77, p = 0.0003); Rad50 mRNA (T(4) = 9.55, p < 0.0001); Rad54l2 mRNA (T(4) = 16.8, p < 0.0001). n = 3/group. Significance assigned by one-tailed t-test, *** p < 0.001, **** p < 0.0001 vs. control RCNs. This experiment was repeated with miR-711 inhibitor and miR-ive inhibitor. qPCR analysis for levels of miR-711, Rad50 and Rad54l2 (D): miR-711 (F(2,6) = 41.13, p = 0.0008 after IR with miR-ve inhibitor, compared to control), Rad50 mRNA (F(2,6) = 79.83, p < 0.0001 after IR with miR-ve inhibitor, compared to control; for miR-711 inhibitor, compared to miR-ve inhibitor, p = 0.0200), Rad54l2 mRNA (F(2,6) = 41.51, p = 0.0003 after IR with miR-ve inhibitor, compared to non-irradiated control; for miR-711 inhibitor, compared to miR-ve inhibitor, p = 0.0016). n = 3/group, *** p < 0.001, **** p < 0.0001 vs. control; ^ p < 0.05, ^^ p < 0.01 vs. corresponding 8Gy + miR-ve inhibitor group.