Fig. 3

The formation of the m6A methyltransferases complex and the increase in total m6A modification levels in chondrocytes are facilitated by circFTO. (A) The potential RBPs were predicted by Circular RNA Interactome and ENCORI. (B) The Circular RNA Interactome predicted the interaction between hsa_circ_0005941 and WTAP, as well as METTL3. (C) ENCORI predicted the interaction between hsa_circ_0005941 and WTAP, as well as METTL3. (D) The interaction between circFTO and WTAP, METTL3, as well as METTL14 was assessed using a pull-down assay. (E) The interaction between circFTO and WTAP, METTL3, as well as METTL14 was evaluated through an RIP assay. (F) IF and FISH assays showing the colocalization of circFTO and WTAP, METTL3, as well as METTL14 in chondrocytes. (G) The level of m6A-modified circFTO was determined using an m6A RIP assay. (H) The expression of circFTO in chondrocytes was detected via qRT-PCR. (I-K) The impact of circFTO on the endogenous interaction between WTAP with METTL3 and METTL14 was examined using a co-IP assay. (L-M) Total m6A modification levels were quantified through an RNA m6A quantification assay. Data are presented as mean ± SD, n = 3; ***p < 0.001, ##p < 0.01, ###p < 0.001