Figure 7.

Investigation of the Functions of RA-Regulated Genes Stra8, Sox30, Agpat3, Wdr91, and Fam57a in Spermatogenesis Using the RA/pSC Model

(A) Quantitative evaluation of mRNAs in feeder-free mSSCs, in which the shRNAs of different genes were expressed from the lentiviral GFP-expressing vector using qRT-PCRs. Error bars indicate mean ± SD from three independent experiments. Asterisks represent statistically significant differences (^∗p < 0.05, ^∗∗p < 0.01).

(B) The growth curves of gene-knockdown mSSC lines on MEF feeders. The data were from three independent experiments. Asterisk denotes statistically significant difference (^∗p < 0.05).

(C) Effects of RNA knockdown on the induction of SYCP3-positive cells using the RA/pSC model. The images were taken on day 6 of induction. The white boxes frame areas where the co-localization of GFP and SYCP3 signals were or were not observed. The ratio of S cells to the total GFP⁺ cells was calculated based on the results from three independent assays (n = 3). The quantification data were normalized by the ratio of S cells in the Sh-CTRL sample, and the mean value for each gene is shown on the right of the immunostaining images. Asterisks denote statistically significant differences (^∗∗∗p < 0.001; n.s., no significance).