Identification of CHD1L as an Important Regulator for Spermatogonial Stem Cell Survival and Self-Renewal

Figure 5

Chd1l is involved in GDNF signaling pathway. (a) Effects of supplementation with higher levels of GDNF on Chd1l gene expression in mouse SSCs. (b) Effects of GDNF withdrawal or replacement on Chd1l gene expression in mouse SSCs. Mouse SSCs were normally maintained in the culture medium supplemented with 15 ng/mL GDNF. GDNF was withdrawn from the culture medium for 24 hours, followed by GDNF restimulation for another 12 hours. Total RNAs were harvested at the indicated time and subjected to quantitative RT-PCR analyses. (c) GDNF-dependent downstream genes are regulated by Chd1l in mouse SSCs. Mouse SSCs were infected with scrambled shRNA or Chd11 shRNA#2 lentivirus. After 48 hours, total RNAs were harvested and subjected to quantitative RT-PCR analyses with the indicated primers. Data presented here are the mean ± SD of three independent experiments. (versus normal SSC culture medium in panels (a) and (b); versus scrambled control shRNA in panel (c)), and (GDNF restimulation versus GDNF withdrawal). SSCs: spermatogonial stem cells; GDNF: glial cell line-derived neurotrophic factor.

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