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. 2014 May 19;9(5):e97909. doi: 10.1371/journal.pone.0097909

Figure 4. Essential role of Sema4D in estrogen-mediated vaginal apoptosis.

Figure 4

(A) Both OVX and OVX-oil treatment of WT (Sema4D+/+) mice result in a significant decrease in cleaved caspase-3 level in vaginal tissue relative to cleaved caspase-3 levels in sham-operated mice (Sham). All vaginal tissue samples were taken from 5-week-old mice. Levels of cleaved caspase-3 were significantly higher in vaginal tissues from WT OVX-E2 mice than in vaginal tissues from WT OVX or WT OVX-oil mice. Levels of cleaved caspase-3 in vaginal tissue did not differ among OVX, OVX-E2, and OVX-oil Sema4D−/− mice; these findings indicate that Sema4D is essential to estrogen-mediated vaginal apoptosis. Each value represents the mean ± SEM of 5 mice. *P<0.05, ANOVA. (B, C) Both TUNEL assays and cleaved caspase-3 immunohistochemistry with vaginal tissue sampled from sham-operated (Sham) 5-week-old WT female (Sema4D+/+) mice show that the number of apoptotic epithelial cells is significantly larger than that in samples from Sham-treated Sema4D−/− mice. OVX-oil treatment of WT mice significantly decreases apoptotic cell number in 5-week-old vaginal epithelia relative to that in Sham; compared with OVX-oil treatment, OVX-E2 treatment of WT mice induces a significant increase in apoptotic cell number in 5-week-old vaginal epithelia, comparable to the level of Sham. OVX-E2 treatment does not induce significant apoptosis in 5-week-old vaginal epithelia of Sema4D−/− mice. Data are shown as means ± SEM; n = 5 per group.