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 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.