Fig. 7

CircCD44 prolonged C-Myc mRNA half-life in an IGF2BP2-dependent manner. A Relative C-Myc RNA level in cells with the indicated modification; ***, p < 0.001. B Immunoblot for C-Myc in cells with the indicated modifications. C mRNA half-life of C-Myc in TNBCs with different modifications; RNA was collected at different times and subjected to qRT–PCR assay, and the relative C-myc RNA was detected and normalized, ***, P < 0.001. D Left: BT549 cells were transfected with IGF2BP2-specific shRNAs, and IGF2BP2 WT/Mut was then transfected to re-express different IGF2BP2 alleles. Right: BT549 cells were transfected with circCD44 WT/Mut. The relative C-Myc RNA level was detected; ***, p < 0.001. E Immunoblot for IGF2BP2 and C-Myc in BT549 cells with the indicated modifications. F Half-life of C-Myc mRNA in BT549 cells with the indicated modifications; ***, P < 0.001. G Immunoblot for IGF2BP2 in cells with circCD44 knockdown or overexpression. H RIP-IB assay. m6A antibody was used for RIP assay, and m6A, C-Myc and IGF2BP2 were measured in cells with the indicated modifications. C-Myc was transfected into all cells to avoid endogenous expression variation. I The RIP assay in (H) was applied, and the complex was subjected to qRT–PCR assay. The relative RNA level of C-Myc was detected, ***, p < 0.001. Data are representative of at least 2–3 experiments with similar results