Fig. 4

IGF2BP2 affected the mRNA stability of PLOD2 by participating in the PLOD2 m⁶A modification. A By using fragmented RNA, MeRIP-qPCR was used to analyze the enrichment of m⁶A in 5′UTR, CDS, or 3′UTR of PLOD2 in control or ALKBH5 overexpression GC-2 cells under oxidative stress. B, C In control and ALKBH5-overexpressing GC-2 cells, the relative luciferase activity of F-Luc/R-Luc of pmirGLO-PLOD2-3'UTR-WT and the relative activity of F-Luc/R-Luc of pmirGLO-3′UTR-MUT were determined. D PLOD2 mRNA was analyzed by RIP-qPCR using IGF2BP2 antibody in control or ALKBH5-overexpressing GC-2 cells. E–G In GC-2 cells transfected with control or siRNA-IGF2BP2 siRNAs, IGF2BP2 and PLOD2 mRNA (E) and protein (F and G) expression levels were detected by RT-qPCR and western blotting. H The mRNA of PLOD2 was determined by RT-qPCR after transfecting GC-2 cells with control siRNA or siRNA-IGF2BP2 for 24 h and then further treating them with Act-D