(A and B) Expression analysis of spermatogonial stem cell markers (Pou5f1, Sox2, and Nanog) (A) and meiotic differentiation markers (Scp3, Dmc1, Spo11, and Stra8) (B) upon prolonged (24-, 48-, 72-, and 96-h) Wnt3A treatment in Gc1-Spg cells. (C) Western blot analysis of Pou5f1 and Stra8 upon prolonged (24, 48, 72, and 96 h) Wnt3A treatment in Gc1-Spg cells. The significance values in panels A and B are represented with respect to 24-h Wnt3A treatment. Data in panels A and B are plotted as means ± SD, n = 4. ***, P ≤ 0.001; **, P ≤ 0.01; *, P ≤ 0.05 (two-tailed Student's t test).