Fig. 5. FMR1 regulates EGFR mRNA stability via m⁶A-dependent manner.

a RT-qPCR was used to test the expression of EGFR when FMR1 was stably overexpressed or knockdown. b RT-qPCR was used to test the stability of EGFR mRNA in the indicated CRC cells. c Prediction of specific binding sites recognized by FMR1 protein on the EGFR mRNA sequence. d Luciferase activity assay showed that FMR1 protein influences the luciferase activity of EGFR mRNA (654 and 797 position) in SW480 and HCT116 cells. e Schematic representation of wild-type (EGFR mRNA-wt) and mutant (EGFR mRNA-mut) EGFR mRNA constructs. f Luciferase activity assay showed that FMR1 protein influences the luciferase activity of EGFR mRNA (3785 position which is locating on the EGFR 3'UTR) in SW480 and HCT116 cells. g Schematic representation of wild-type (EGFR mRNA-wt) and mutant (EGFR mRNA-mut) EGFR mRNA constructs. h The binding between FMR1 protein and EGFR mRNA (3785 position) was validated by RNA EMSA. i RIP-derived RNA in HCT116 cells were measured by RT-qPCR. j Gene-specific m⁶A qPCR validation of m⁶A levels of EGFR mRNA (3785 position) in HCT116 cells. *P < 0.05,**P < 0.01, ***P < 0.001, ****P < 0.0001. NS not significant.