Figure 1
Expression of ZMYM3 in cultured mouse spermatogonial stem cells and in cells of the mouse testis. (a) Zmym3 mRNAs in cultured mouse SSCs are down-regulated by GDNF (n=3). (b) Expression of ZMYM3 in cultured SSCs. Note that the signal of ZMYM3 is stronger in SSCs which are weaker in GFRα1 staining (arrow heads) but weaker in SSCs which are stronger in GFRα1 staining (arrows). (c–e) Zmym3 mRNAs and proteins in cultured SSCs are up-regulated by RA as determined by qRT-PCR (c) (n=3) and Western blot assays (d) (n=4). (e) is the quantitative densitometry of the Western blot result in (d). (f) Western blot of ZMYM3 in cytoplasm and nuclear fractions of cultured SSCs. Total: total cell lysates; Cyt: cytoplasmic fraction; Nuc: nuclear fraction; MVH: Mouse Vasa Homolog, a germ cell specific marker and a cytoplasmic protein; LMNB1: a component of the nuclear lamina and a nuclear protein. (g) Immunohistochemical analysis of ZMYM3 distribution in different testicular cells of adult mice. Note that a small dot in the spermatocyte most likely representing the sex body was positively immunostained. However, it turned out to be a nonspecific signal because it was also observed in Zmym3 KO mice (Figure 2e). Roman numerals indicate the stage of spermatogenesis. SG-A, spermatogonia type A; plpSC, preleptotene spermatocytes; lepSC, leptotene spermatocytes; zygSC, zygotene spermatocytes; pacSC, pachytene spermatocytes; rST, round spermatids; eST, elongating spermatids; SE, Sertoli cells; LE, Leydig cells. (h–j) Whole-mount co-immunostaining of ZMYM3 with PLZF and GFRα1 on seminiferous tubules from P6 mice and c-KIT on those from P8 mice
