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. 2015 Jun 29;24(18):5234–5249. doi: 10.1093/hmg/ddv244

Figure 3.

Figure 3.

FANCB regulates the maintenance of undifferentiated spermatogonia. (A) Immunostaining of testicular sections at 10 weeks old. WT1 is a marker for Sertoli cells. Dotted circles are the areas of representative tubules. (B) Quantification of seminiferous tubule areas (103 μm2) at 4 and 10 weeks after birth. Mean and SEM for independent mice are shown. A total of 100 tubules from three independent mice were analyzed at 4 and 10 weeks, for both wild-type and Fancb mutant mice. (C) TUNEL assays of testicular sections at 10 weeks old. (D) Scoring of the number of TUNEL-positive cells per germ cell-containing tubule. Mean and SEM for independent mice are shown. Total numbers of tubules analyzed at 4 and 10 weeks, respectively: 303 and 430 wild-type, and 394 and 138 Fancb mutant mice. Three independent mice were analyzed for each dataset. (E and F) Immunostaining of testicular sections at 10 weeks old for panels in E and at 6-months old for panels in F. PLZF is a marker for undifferentiated spermatogonia. Arrowheads: PLZF-positive cells. (G) Scoring of the percentage of PLZF-positive tubules per total tubule. One hundred tubules from three independent mice were analyzed at 4 weeks, 10 weeks and 6 months, in both wild-type and Fancb mutant mice. Mean and SEM for independent mice are shown. (H) Scoring of the number of PLZF-positive cells per PLZF-positive tubule. One hundred tubules from three independent mice were analyzed at 4 weeks, 10 weeks and 6 months, both for wild-type and Fancb mutant mice. Mean and SEM for independent mice are shown. (I and J) Immunostaining of testicular sections at 10 weeks old for panels in I, and at 6 months old for panels in J. (K) Scoring of the percentage of γH2AX-positive tubules per total tubule. Mean and SEM for independent mice are shown. One hundred tubules from three independent mice were analyzed at 4 and 10 weeks, in both wild-type and Fancb mutant mice. One hundred tubules (two independent mice) were analyzed at 6 months, in both wild-type and Fancb mutant mice. Scale bars: 100 µm. *P < 0.05. Unpaired t-test.