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 7. Influence of CASC7 silencing on level of MiRNA-122-5p and CBL as well as identification and verification of the direct binding of MiRNA-122-5p and CASC7 in human SSC line. (A) Real-time qPCR showed changes of CASC7 by CASC7 siRNA 1, CASC7 siRNA 2, and CASC7 siRNA 3 in human SSC line. * indicated statistically significant differences (P< 0.05) of CASC7 siRNA 1-, 2-, and 3-treated human SSC line compared with the control siRNA. (B) Real-time qPCR demonstrated miRNA-122-5p expression changes in human SSC line at day 2 after transfection of CASC7 siRNA 2 and the control siRNA.* denoted statistically significant differences (p<0.05) between CASC7 siRNA 2-treated cells and the control siRNA.(C) Real-time qPCR showed the change of CBL in human SSC line at day 2 after transfection of CASC7 siRNA 2 and the control siRNA.* showed statistically significant differences (p< 0.05) between CASC7 siRNA 2-treated cells and the control siRNA. (D) Validation of the direct binding of miRNA-122-5p to wild type CASC7 and the mutated CASC7 by dual luciferase reporter assays. * indicated statistically significant differences (p< 0.05) in human SSC line between miRNA-122-5p mimics and miRNA mimics control. (E) The sequences of miRNA-122-5p complement with CASC7.