LRP6 Knockdown Attenuated the Effects of miR-202-3p Inhibition on Sertoli Cells
(A) CCK-8 assay displayed the proliferation of human Sertoli cells treated with miRNA inhibitor control, miR-202-3p inhibitor, and miR-202-3p inhibitor+LRP6 siRNA-3 for 5 days. (B) Western blot showed the expression of PCNA in Sertoli cells treated with miRNA inhibitor control, miR-202-3p inhibitor, and miR-202-3p inhibitor+LRP6 siRNA-3 at 48 hr. Results of three independent assays were concluded in (C). (D) Western blot showed the expression of GDNF, SCF, BMP4, FGF2, and CXCL12 in Sertoli cells treated with miRNA inhibitor control, miR-202-3p inhibitor, and miR-202-3p inhibitor+LRP6 siRNA-3 at 48 hr. Results of three independent assays were concluded in (E). (F) Annexin V and propidium iodide (PI) staining and flow cytometry showed the percentage of apoptosis in human Sertoli cells treated with miRNA inhibitor control, miR-202-3p inhibitor, and miR-202-3p inhibitor+LRP6 siRNA-3 at 48 hr. Results of three independent assays were concluded in (G). A total of 15,000 cells were analyzed. (H) Western blots demonstrated cPARP, Bcl2, and Bax proteins in human Sertoli cells treated with miRNA inhibitor control, miR-202-3p inhibitor, and miR-202-3p inhibitor+LRP6 siRNA-3 at 48 hr. Results of three independent assays were concluded in (I). *p < 0.05; **p < 0.01.