Research Paper Volume 15, Issue 5 pp 1564—1590

Figure 2. Knockdown of circRBMS3 inhibits the migration and invasion of OS cell lines in vitro. (A, B) The expression levels of circRBMS3 and RBMS3 mRNA in HOS and 143B cells after stable transfection of circRBMS3 short hairpin RNAs or vector plasmids were detected by qPCR. Data represent the mean ± SD (n = 3). *P < 0.05. (C) SiRNA-mediated circRBMS3 knockdown suppressed OS cell proliferation, as determined by CCK-8 assays. Data represent the mean ± SD (n = 6). (D) CircRBMS3 knockdown suppresses cell growth, as determined by colony formation assays (details are shown in the insets). Error bars represent the mean ± SD of three independent experiments. * P < 0.05. (E) HOS and 143B cells were transfected with sicircRBMS3, followed by Annexin V-FITC/PI staining. The percentage of apoptotic cells is shown as the mean ± SD from three independent experiments. * P < 0.05, significantly different compared with the vector group. (F) The effect of sicircRBMS3 on cell migration capability was evaluated by wound healing assays using HOS and 143B cells. Data are the mean ± SD, n = 3. *P < 0.05. Scale bar, 200 μm. (G) CircRBMS3 knockdown suppresses cell migration and invasion abilities of HOS and 143B cells, as evaluated by transwell migration and Matrigel™ invasion assays. Data represent the mean ± SD (n = 3). * P < 0.05. Scale bar, 200 μm. (H) CircRBMS3 knockdown did not affect linear RBMS3 expression. (I) Total cell lysates were separated by SDS-PAGE and Coomassie blue staining. (J) Cell lysates were precipitated with anti-RBMS3 antibody followed by SDS-PAGE and Coomassie blue staining. Transfection with circRBMS3 siRNA did not affect the interaction of RBMS3 with its partners.