Gene and protein characterization of the freshly isolated human Sertoli cells. (A) RT-PCR showed the expression of numerous genes, including WT1, GATA4, GATA1, GDNF, BMP4, SCF, FGF2, EGF, FHSR, AR and ABP. ACTB was used as a loading control, and RNA without reverse transcriptase enzyme but with PCR of ACTB primers was used as a negative control (NC). (B-I) Immunofluorescence revealed the expression of WT1 (B), GDNF (C), SCF (D), BMP4 (E), VIM (F), PCNA and GATA4 (G), SMA (H), and CYP11A1 (I) in isolated human Sertoli cells. Scale bars in B, C, D, F, H =50 μm; scale bars in E, G, I =20 μm.