Fig. 3

Identification of spermatogonia isolated from 7-day testes by magnetic-activated cell sorting. a, b The purity of the isolated spermatogonia were identified by flow cytometry analysis, showed that spermatogonia with high purity were isolated by magnetic antibodies. Error bars represent SD (n = 5). **p < 0.01. c Identification of the isolated spermatogonia by quantitative PCR, showed high levels of Plzf and Gfra1 in Thy1 positive cells and high levels of TFEB, Kit and Sohlh2 in c-Kit positive cells. Error bars represent SD (n = 5). **p < 0.01. d Western blot detection of TFEB in testis, Sertoli cells and isolated spermatogonia, indicated it was mainly expressed in c-Kit positive spermatogonia. e Immunofluorescence analysis showed the high expression of TFEB (green fluorescence) in c-Kit positive spermatogonia. Bar: 20 μm