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. 2020 Nov 11;11(11):965. doi: 10.1038/s41419-020-03183-5

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© The Author(s) 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 5 — A Confocal images of TUNEL assay examining control and quercetin-treated oocytes. The green fluorescence indicates TUNEL-positive oocytes. Scale bar, 25 μm. B Confocal images showing levels of activated Caspase 3 in control and quercetin-treated oocytes. Scale bar, 25 μm. C Confocal images showing autophagosomes (LC3-II puncta) in control and quercetin-treated oocytes. Scale bar, 25 μm. D Confocal images showing SOD2K68c/SOD2 in young and aged oocytes. Scale bar, 25 μm. E Western blot analysis showed reduced SIRT3 expression in GV stage oocytes from aged mice compared with young mice. β-action served as an internal control. F Schematic illustration of the experimental protocol to determine whether quercetin treatment reduces ROS accumulation levels and increases the extent of SOD2K68 acetylation by Sirt3. G Quercetin lowers the acetylation levels of SOD2K68 acetylation and ROS upon the reduction of SIRT3 expression. Scale bar, 25 μm. H The percentage of TUNEL-positive oocytes in control and quercetin-treated oocytes. I The Caspase3 was determined by immunofluorescence in control and quercetin-treated oocytes. J Quantification of LC3 intensity in control and quercetin-treated oocytes. K The levels of SOD2K68 are negatively associated with SOD2 enzymatic activity in M II stage oocytes from aged mice and young mice. L Western blot analysis showed the reduced Sirt3 expression in oocytes from aged mice compared with young mice. M Quantitative analysis of fluorescence intensity showing the acetylation levels of SOD2K68 after injecting Control, siSirt3, and siSirt3 + Quercetin treatment (Que). N Quantitative analysis of ROS fluorescence intensity after injecting Control, siSirt3, and siSirt3 + Quercetin treatment (Que). DNA was stained with DAPI. Data are means ± SD of at least three independent experiments. Differences between two groups were analyzed by two-tailed unpaired Student’s t-tests. Multiple comparisons between more than two groups were analyzed by one-way ANOVA test. *P < 0.05 vs. control; **P < 0.01 vs. control as calculated by two-tailed unpaired Student’s t-tests.