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. 2022 Jul 27;19(2):644–659. doi: 10.1080/15548627.2022.2094671

Figure 3.

Figure 3.

Epg5 knockout results in GCs aging and cell death. (a) Histoenzymatic detection of GLB1/β-gal. The blue indigo precipitates (resulting from X-Gal lysis by GLB1/β-gal) are evident in the GCs of 12-week-old epg5 knockout mice. The red asterisks indicate follicles positive for GLB1/β-gal signals. Scale bar: 200 µm. (b) Quantification of GLB1/β-gal-positive GCs in ovary sections (n = 5 for each of the indicated groups). (C) TUNEL assay to evaluated GC apoptosis in 12-week-old epg5 knockout and control ovaries. The white asterisks indicate follicles positive for TUNEL. Scale bars: 200 µm, 20 µm. (g) Quantitative analysis of TUNEL-positive cells of ovary sections in C (n = 4 for each group). (d) PCNA immunofluorescence assays and (h) the quantification of PCNA positive granulosa cells in epg5 knockout and control ovaries are shown (n = 4 for each group). Scale bars, 20 µm. (e) CASP3 Immunofluorescence analysis of epg5 knockout and control ovaries sections. Scale bars: 50 µm, 20 µm. (i) Quantitative analysis of CASP3-positive cells of ovary sections in E (n = 3 for each group). (f) γ-H2AX immunofluorescence assays and J the quantification of γ-H2AX positive GCs in epg5 knockout and control ovaries are shown (n = 5 for each group). Scale bars: 20 µm. Data are expressed as the mean ± S.E.M.