Research Paper Volume 15, Issue 5 pp 1564—1590

Figure 8. circRBMS3 functions as a miR-424 sponge to promote tumorigenesis in vivo. (A, B) Nude mice were injected with 5 × 10⁶ 143B stable cells. Four weeks later, tumors were dissected and photographed. (C) The graph represents tumor volumes (v = ab²/2) at injection with control cells or cells transfected with circRBMS3 short hairpin (sh)-RNA or co-transfected with circRBMS3 shRNA and miR-424 sponge (n = 6 per group). Data represent the mean ± SD (n = 8). (D) Average tumor weight in each group at the end of the experiment. Data represent the mean ± SD (n = 8). * P < 0.05. (E) In vivo imaging of tibia tumor. (F) Average radiance of orthotopic xenograft nude mice. (G) The limb weight of orthotopic xenograft nude mice. (H) RT-qPCR analysis of EIF4B and YRDC expression in tumors from xenograft mice. (I, J) Knockdown efficiency of circRBMS3 and miR-424 in tumors from orthotopic xenograft nude mice. (K) Western blot analysis of EIF4B and YRDC in tumors from xenograft mice. (L) Histological analysis of tumor tissues by hematoxylin and eosin staining. EIF4B and YRDC expression was examined by immunohistochemistry. Representative images are shown. (M) MicroCT quantification of the specific trabecular bone volume [BV/TV (%)] and the cortical BV (mm³)] were calculated for the tibia of tumor-bearing mice in the different groups. (N) Schematic illustration of the circRBMS3/miR-424 axis.