Research Paper Volume 12, Issue 21 pp 21971—21991

CircGLCE alleviates intervertebral disc degeneration by regulating apoptosis and matrix degradation through the targeting of miR-587/STAP1

Figure 2. Knockdown of CircGLCE induces apoptosis and the upregulation of matrix-degrading enzymes in NP cells. (A) Forty-eight hours after transfecting NP cells with CircGLCE siRNA or a negative control, RT-qPCR showed that CircGLCE expression was effectively downregulated by silencing (n=6, ***P < 0.001, unpaired two-tailed Student’s t test). (B) Flow cytometry showed an increased rate of apoptosis in NP cells in response to silencing CircGLCE, which was indicated by increased accumulation of signals in Q2 (n=3, *** P<0.001, unpaired two-tailed Student’s t test). (C) NP cells were transfected with CircGLCE lentivirus or the corresponding negative control, and RT-qPCR showed that CircGLCE expression was effectively upregulated (n=6, ***P < 0.001, unpaired two-tailed Student’s t test). (D) Flow cytometry showed that overexpressed CircGLCE partially reversed the apoptosis of NP cells that was promoted by treatment with IL-1β (n=3, *** P<0.001, unpaired two-tailed Student’s t test). (E) Shown by immunofluorescence analyses, silencing CircGLCE resulted in increased expression of MMP13 and ADAMTS 5 as well as decreased expression of COL II and Aggrecan. (F) Western blot analysis (n=3) confirmed the results in (E). Data are the mean #x00B1; SEM.