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. 2020 Nov 20;12(24):25207–25228. doi: 10.18632/aging.104123

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Copyright: © 2020 Hua et al.

This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

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Effects of Pol β knockdown on oocytes apoptosis. (A) Western blotting analysis indicates that Pol β siRNA group have barely expression. (B) Bar graphs show a result with Annexin V/PI staining. Statistics of oocytes number per non-Injctrl/ Scramble siRNA/ Pol β siRNA oocytes including Annexin V(-)/PI(-), AnnexinV(+)/PI(+) and Annexin V(+)/PI(-). Bar graphs show Pol β siRNA group more Annexin V(+)/PI(-), more Annexin V(+)/PI(+), and less Annexin V(-)/PI(-) compared to non Injctrl/ Scramble siRNA group, (n=4 per group; n stands for the number of mice, ***P<0.001; ***P<0.001; **P<0.01, respectively). (C) Impact of siRNA silencing of Pol β on genomic integrity and survival of mouse oocytes in response to genotoxic stress. In response to H₂O₂ treatment (250 μM), AC3 levels were higher (n stands for the number of oocytes, **P < 0.01, Student’s t test). Photomicrographs are representative of the AC3 levels in the siRNA-silenced oocytes: scramble, Pol β (green). Arrowheads show the nuclear region. Oocytes are counterstained with DAPI (blue). (D) Survival was lower in the oocytes in which the expression of Pol β had been silenced compared to those in controls (scramble siRNA) (n stands for the number of mice, ***P < 0.001). (E) Scatter graphs represent the 8-OHdG in oocytes, with a significant increase in Pol β siRNA oocytes compared to that in non-Injctrl/ scramble siRNA oocytes (n=6 per group; n stands for the number of mice, **P<0.01). (F) Scatter graphs represent the relative ratio of AP sites in oocytes, with a significant increase in Pol β siRNA oocytes compared to that in non-Injctrl/ scramble siRNA oocytes (n stands for the number of mice, ***P<0.001, Student’s t test). (G) Scatter graphs represent the percentage of positive comet tail in oocytes, with a significant increase percentage of positive comet tail in Pol β siRNA oocytes compared to that in non-Injctrl/ scramble siRNA oocytes (n stands for the number of mice, **P<0.01, Student’s t test). Each experiment was repeated three times. All graphs show the means ±SD.