Fig. 5

WTAP mediated miR-92b-5p maturation in an m6A-dependent manner. A The heatmap showed that differential miRNAs expression profiles obtained by miRNA-Seq analysis between the shNC, LPS + shNC, and LPS + shWTAP groups. B The sequence of pri-miR-92b and the potential m6A site of pri-miR-92b. Green indicated miR-92b-5p sequence, blue indicated pre-miR-92b sequence, and red indicated m6A sites. C Chondrocyte was infected with shWTAP, shNC, mock, or WTAP, and then dealt with 40 ng/mL LPS for 48 h. The qRT-PCR was used to assess the RNA expression of pri-miR-92b, pre-miR-92b, and miR-92b-5p. D MeRIP coupled with qRT-PCR was used to detect the m6A levels of pri-miR-92b in chondrocyte with or without LPS. E Chondrocyte was infected with shWTAP, shNC, mock, or WTAP, and then the m6A levels of pri-miR-92b were evaluated using MeRIP coupled with qRT-PCR. F Relative luciferase activity of the wild-type or mutant-type of [m6A] pri-miR-92b reporter vectors was assessed in WTAP-overexpression chondrocyte. G Quantification of miR-92b-5p, pre-miR-92b, and pri-miR-92b in the reaction mixture was detected by qRT-PCR, and found that mutation of [m6A] pri-miR-92b abolished pri-miR-92b processing in the in vitro reaction system. LPS, lipopolysaccharide; WTAP, WTAP overexpression adenovirus; mock, negative control corresponding to WTAP; shWTAP, WTAP knockdown adenovirus; shNC, negative control corresponding to shWTAP. N = 3. ^n.s.P > 0.05, ^**P < 0.01, and ^***P < 0.001