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. 2018 Oct 25;14:1–19. doi: 10.1016/j.omtn.2018.10.012

Figure 2.

Figure 2

Differentially Expressed miR-202-3p Inhibits the Proliferation of Human Sertoli Cells

(A) Real-time qPCR revealed the expression of miR-202-3p in both OA and SCOS Sertoli cells (n = 20). (B and C) CCK-8 assay showed the growth curve of human Sertoli cells for 5 days in the pre-miR group (B) and the pre-miR inhibitor group (C) after virus infection and puromycin screening. (D) EDU incorporation assay showed the EDU-positive cells in human Sertoli cells after virus infection and puromycin screening. Cell nuclei were counterstained with Hoechst 33342. The percentages of EDU-positive cells were counted out of 500 total cells from three independent experiments. (E) Immunofluorescence revealed the ki-67-positive cells in the four cell strains. Cell nuclei were counterstained with DAPI. The percentages of ki-67-positive cells were counted out of 500 total cells from three independent experiments. (F) Western blots demonstrated the expression of PCNA and cell-cycle proteins in human Sertoli cells at 72 hr after virus infection and puromycin screening. β-actin served as a loading control of proteins. Results of the pre-miR group were normalized to the Normal ctrl group, and results of the pre-miR inhibitor group were normalized to the inhibitor ctrl group. *p < 0.05; **p < 0.01; ***p < 0.001. Scale bars, 10 μm (D and E).