Additional file 1: of Expression of transcriptional factor EB (TFEB) in differentiating spermatogonia potentially promotes cell migration in mouse seminiferous epithelium

Figure S1. Isolation and culture of spermatogonia from 7-day testes. (a-b) Enzymatic digestion of seminiferous tubules into fragments and single cells. (c) Testicular single cells were further performed differential attachment, showed the somatic cells present as triangle- or spindle-liked forms were attached to the plate, whereas the round unattached cells were spermatogonia. (d) Spermatogonia purified by magnetic-activated cell sorting. (e) Culture of the somatic cells isolated by differential attachment, containing Sertoli cells and myoid cells. (f) Culture of the Thy1 positive spermatogonia. Bar: 200 μm in (a), (b); 100 μm in (c)-(e); 50 μm in (f). Figure S2. Immunofluorescent staining of GFRA1 in cultured spermatogonia. The result showed most cells were positive for GFRA1, suggesting high purity of undifferentiated spermatogonia. Bars: 20 μm. Figure S3. Immunofluorescent staining of TFEB in GC-1 cells. The result showed that RA treatment induced TFEB nucleus translocation. Bars: 20 μm. Table S1. Oligonucleotide primer sequences used for qRT-PCR. (DOCX 3401 kb