Fig. 6

Effect of chd4 deletion in Dmrt1 and Plzf expression. A Histological sections of wild type and ddx4-chd4^−/− testis from 1, 4, and 7 dpp mice stained with Chd4 and Dmrt1 antibodies. Arrows indicate examples of Chd4 positive spermatogonia (Wt) or Chd4 negative staining in ddx4-chd4^−/− spermatogonia and the correspondent cell stained for Dmrt1. B Quantitation of fluorescence intensity of Dmrt1 expression on spermatogonia is shown for wild type (Wt) and ddx4-chd4^−/− knockout at ages of 1 dpp (Wt = 82.17 ± 22.02, n = 272; and KO = 37.20 ± 10.01, n = 287, P < 0.0001, Student t test) and 4 dpp (Wt = 71.09 ± 20.93, n = 242; and KO = 43.47 ± 14.88, n = 198, P < 0.0001, Student t test). Values represent the median fluorescence intensity ± standard deviation, n = total number of cells analyzed from 3 biological replicates using 3 different mice. C Histological sections of wild type and ddx4-chd4^−/− testis from 1, 4, and 7 dpp mice stained with antibodies against Plzf. D Quantitation of fluorescence intensity of Plzf expression on spermatogonia is shown for wild type and ddx4-chd4^−/− knockout at ages of 1 dpp (wt = 17.94 ± 4.10, n = 300; and ddx4-chd4^−/−  = 10.89 ± 4.07, n = 339, P < 0.0001, Student t test) and 4 dpp (Wt = 17.78 ± 4.70, n = 282; and ddx4-chd4^−/− = 9.11 ± 3.75, n = 230, P < 0.0001, Student t test). Values represent the median fluorescence intensity ± standard deviation, n = total number of cells analyzed from 3 biological replicates using 3 different mice. E Diagram representing Chd4, Dmrt1, and Plzf relationship in spermatogonia stem cell maintenance. Note that our studies do not address the possibility that Plzf may be a direct target of CHD4