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. 2022 Apr 19;13(4):377. doi: 10.1038/s41419-022-04837-2

Fig. 1. UHRF1 displays a temporal and spatial expression pattern in SCs of both humans and mice.

Fig. 1

A Double immunostaining with anti-UHRF1 and anti-WT1 antibodies on wild-type (WT) mouse testicular sections at embryonic day 14.5 (E14.5) and postnatal day 70 (P70) and human testis sections at fetal 20 weeks and adults are shown. Scale bar = 50μm. B, C Co-immunofluorescent staining for UHRF1 and WT1 (a SC marker) on WT mouse testis sections at E14.5, E16.5, E18.5, P1, P3, P5, P7, P10, P14, and P21 are shown. Nuclei were stained with DAPI. Scale bar = 50 μm. D The quantifications of the ratio of both UHRF1+ and WT1+ cells to WT1+ cells per tubule for B and C are shown. Data are shown the mean values ± SEM. For E14.5, E16.5, and E18.5, n = 3 mice; For P1, P3, P5, P7, and P10, n = 5 mice; For P14, P21, and P70, n = 7 mice. Approximately 20–30 round seminiferous tubules per mouse were randomly chosen to count. ***P < 0.001 by one-way ANOVA. E Double immunostaining with anti-UHRF1 and anti-WT1 on isolated primary SCs from P3 and P21 WT testes are shown. Scale bar = 20μm. Right histogram shows the percentage of both UHRF1+ and WT1+ cells to WT1+ cells. Data show mean values ± SEM, n = 3 mice. Approximately 30 round seminiferous tubules per mouse were randomly chosen to count. F Double immunostaining with UHRF1 and WT1 on primary SCs from P21 testes cultured with or without stem cell differentiation inhibitors (1 μM PD0325901 + 10 μM SP600125 + 10 μM SB203580) and 10% FBS for 24 h are shown. Scale bar = 20 μm. G, H Quantification of the percentage of UHRF1-positive cells (G) and cell viability (H) are shown. ns not significant. ***P < 0.001 by unpaired Student’s t-test.