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

Figure 6.

Figure 6

miR-711 inhibition attenuates IR-induced neuronal apoptosis. Western blot for Puma, cleaved caspase-3, PARP, α-fodrin and β-actin (A). (B) Quantification of levels of Puma (F(10,22) = 3.48, p = 0.0007 at 1 h, p < 0.0001 at 3 and 6 h after IR with miR-ve inhibitor, compared to control; for miR-711 inhibitor, compared to miR-ve inhibitor, p = 0.0312 at 30 min, p < 0.0001at 6 h), cleaved caspase-3 (F(10,22) = 5.25, p < 0.0001 at 6 and 24 h after IR with miR-ve inhibitor, compared to control; for miR-711 inhibitor, compared to miR-ve inhibitor, p = 0.0005 at 6, p < 0.0001 at 24 h), cleaved PARP (89kDa) (F(10,22) = 296.3, p = 0.0180 at 3, p < 0.0001 at 6, p = 0.0047 at 24 h after IR with miR-ve inhibitor, compared to non-irradiated control for miR-711 inhibitor, compared to miR-ve inhibitor, p = 0.0225 at 3, p < 0.0001 at 6 h), α-fodrin (120kDa) (F(10,22) = 132.9, p < 0.0001 at 6 and 24 h after IR with miR-ve inhibitor, compared to control for miR-711 inhibitor, compared to miR-ve inhibitor, p = 0.0068 at 6 h, p < 0.0001 at 24 h). n = 3/group. * 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. qPCR quantification (C) of PUMA mRNA (F(10,43) = 351.7, p < 0.0001 at all-time points after IR with miR-ve inhibitor, compared to control; for miR-711 inhibitor, compared to miR-ve inhibitor, p < 0.0001 for 3 h and 6 h), Noxa mRNA (F(10,43) = 389.8, p = 0.0254 at 1, p < 0.0001 at 3 and 6, p = 0.0470 at 24 h after IR with miR-ve inhibitor, compared to control for miR-711 inhibitor, compared to miR-ve inhibitor, p < 0.0001 for 3 and 6 h) and Bim mRNA (F(10,43) = 85.04, p < 0.0001 at 3 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 6 and 24 h). n = 4 for control, n = 5/group for all other groups, * p < 0.05, **** p < 0.0001 vs. control; ^^^^ p < 0.0001 vs. corresponding 8Gy + miR-ve inhibitor group. Cytosolic fractions were used for Western blot for AIF-1, cytochrome c and GAPDH (D). Levels of AIF-1(E)(F(2,9) = 294.1, p < 0.0001 for all comparisons) and Cytochrome c (F)(F(2,9) = 4.62, p < 0.0001 for non-irradiated control vs. miR-ve inhibitor group and for miR-ve inhibitor vs. miR-711 inhibitor group). n = 3/group, **** p < 0.0001 vs. control RCNs; ^^^^ p < 0.0001 vs. 8Gy + miR-ve inhibitor group. (G) LDH was measured 24 h and 48 h after irradiation)(F(5,30) = 131.3, p < 0.0001 at 24 h and 48 after IR with miR-ve inhibitor, compared to non-irradiated control for miR-711 inhibitor, compared to miR-ve inhibitor, p = 0.0002 at 24 h, p < 0.0001 at 48 h; For 24 h vs. 48 h 8gy + miR-ve inhibitor, p < 0.0001; For 24 h vs. 48 h 8Gy + miR-711 inhibitor, p = 0.0045). n = 3/group. **** p < 0.0001 vs. control RCNs; ^^^ p < 0.001, ^^^^ p < 0.0001 vs. corresponding 8Gy + miR-ve inhibitor group; && p < 0.01, &&&& p < 0.0001 vs. equivalent treatment 24 h after 8Gy.