Research Paper Volume 12, Issue 24 pp 25528—25546

miRNA-122-5p stimulates the proliferation and DNA synthesis and inhibits the early apoptosis of human spermatogonial stem cells by targeting CBL and competing with lncRNA CASC7

Figure 2. Effect of overexpression of MiRNA-122-5p on the proliferation, DNA synthesis and early apoptosis of human SSC line. (A) CCK-8 assays revealed the growth curve of human SSC line treated with miRNA-122-5p vector and control lentivirus in 5 days. (B) Western blots demonstrated PCNA expression in human SSC line of miRNA-122-5p vector and control lentivirus. (C) The relative expression of PCNA in human SSC line of miRNA-122-5p vector and control lentivirus after normalization to the signals of their loading control ACTB. (D) EDU incorporation assay showed the EDU-positive cells in human SSC line of miRNA-122-5p vector and control lentivirus. Scale bars =50 μm. (E) Qualification of EDU-positive cells in human SSC line of miRNA-122-5p vector and control lentivirus. (F, G) APC Annexin V and PI staining and flow cytometry showed the percentages of early apoptosis in human SSC line infected with ZsGreen1-labeled miRNA-122-5p lentivirus and control lentivirus. * denoted statistically significant differences (p< 0.05) in human SSC line between control lentivirus and miRNA-122-5p vector.