Figure 3. SM accelerates differentiation of undifferentiated Kit-negative spermatogonia.

A) FACS analysis of total germ cells population from testes of 4dpp mice before (left panel) and after (right panel) removal of Kit positive cells. B) Western blot analysis of Kit, Stra8 and Scp3 expression in undifferentiated Kit negative cells at T0 and after 24 h of culture under gravity or SM. C) Lack of Scp3 organization on nuclear spreads of undifferentiated spermatogonia after 24 h of culture under either unit gravity or SM conditions. D) Scp3 organization on nuclear spreads of total spermatogonia from 4dpp mice, cultured for 24 h under SM or gravity conditions. Preleptotene-like cells are evident only in microgravity samples. E) Histogram representing the percentage of preleptotene-like nuclei in gravity and SM cultures. The values were obtained by counting 300 nuclei in each sample. Data are means ± SD of at least three independent determinations. P≤0.05. F) Representative semiquantitative RT-PCR (left panel) and western blot (right panel) for Scp3 expression in total spermatogonia from 4pp testes, cultured under SM compared to control cells. Similar results were obtained in three separate experiments.