Figure 4

Expression and siRNA knockdown of LIN28 in cultured spermatogonia highly enriched for spermatogonial stem cells (SSCs). (A) Immunostaining of SSCs with anti-LIN28 and anti-PLZF or anti-GFRA1 antibodies. Scale bar, 50 μm. (B) Quantitative PCR measurement of Lin28 mRNA levels (n = 3, mean ± SE) in SSCs after siRNA treatment for 30 hours. (C) Decreased LIN28 protein abundance (43% compared to the control) in SSCs after 30 hours of siRNA treatment. The control SSCs were not treated with Lin28 siRNA. Feeder cells served as a negative control. β-actin served as a loading control. (D) The number of SSCs (n = 3, mean ± SE) with and without Lin28 siRNA treatment. (E) Quantitative measurement of mature let-7g miRNA levels (n = 3, mean ± SE) in SSCs after siRNA treatment for 30 hours.