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

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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.