Fig. 4

A Relative circNHS levels in nuclear and cytosolic fractions of C4-2-IRR cells and 22Rv1 cells are shown. U6 was used as the nuclear control. GAPDH was used as the cytosolic control. B RNA-FISH assay results indicated the location of circNHS in C4-2-IRR cells and 22Rv1 cells. C RIP experiments were performed using an antibody against AGO2 on extracts from C4-2-IRR cells and 22Rv1 cells. D The circRIP was performed in C4-2-IRR cells using a circNHS-specific probe and control probe. The enrichment of miRNAs was detected by RT-qPCR and normalized to the control probe. E Mutation of miR-512-5p binding sites abolished the effects of circNHS on radiosensitivity, as revealed using colony formation assay in C4-2 cells. F WB analysis of XRCC5 and SPRTN protein levels after shcircNHS in C4-2-IRR cells and oecircNHS in C4-2 cells. G WB assays: effects of shcircNHS and/or miR-512-5p inhibitor on XRCC5 protein levels in C4-2-IRR cells (left) and effects of oecircNHS and/or miR-512-5p mimics on XRCC5 protein levels in C4-2 cells (right). H shARv7 and/or oeXRCC5 effects on C4-2-IRR cells survival after IR using clonogenic assay. I The oeARv7 and/or shXRCC5 effects on C4-2 cell survival after IR using clonogenic assay. *P < 0.05, ^**P < 0.01, and ^***P < 0.001 compared with the controls. N.S., not significant. Data are presented as mean ± SEM