Fig 1: METTL3 interacted with RAC2 by m6A methylated modification. A RMBase v3.0 website prediction for m6A methylated modification of RAC2. B RBP suite analysis for METTL3 and RAC2. C–F RT-qPCR and western blot for METTL3 expression analysis in RA tissues C–D and TNF‐α-treated MH7A cells E–F. G RAC2 protein detection by western blot after shNC or shMETTL3 transfection in TNF‐α-treated MH7A cells. H RAC2 mRNA expression was tested via RT-qPCR after actinomycin D treatment and shMETTL3 or shNC transfection. I–G RIP I and MeRIP J were performed to confirm the interaction between METTL3 and RAC2. *P < 0.05, **P < 0.01, ***P < 0.001
Fig 2: Graphical abstract. METTL3 up-regulated RAC2 expression via m6A methylated modification to promote proliferation, oxidative stress and inflammation in TNF-α-stimulated MH7A cells
Fig 3: Overexpression of RAC2 rescued the function of METTL3 knockdown in cell viability and motility. Five groups (con, TNF‐α, TNF‐α + shNC, TNF‐α + shMETTL3, TNF‐α + shMETTL3 + oeRAC2) were divided in MH7A cells. A–B RT-qPCR and western blot for RAC2 expression. C–D CCK-8 assay for cell proliferation and flow cytometry for cell apoptosis. E–F Wound healing assay for cell migration and transwell assay for cell invasion. *P < 0.05, **P < 0.01, ***P < 0.001
Fig 4: Downregulation of RAC2 promoted apoptosis and suppressed viability, migration, invasion in TNF‐α-induced MH7A cells. Four groups (con, TNF‐α, TNF‐α + shNC, TNF‐α + shRAC2) were divided in MH7A cells. A–B RAC2 mRNA and protein detection through RT-qPCR and western blot. C–D Cell proliferation and apoptosis examination by CCK-8 assay and flow cytometry, respectively. E–F Cell migration and invasion analysis by wound healing assay and transwell assay, respectively. *P < 0.05, **P < 0.01, ***P < 0.001
Fig 5: RAC2 overexpression inhibited the effects of METTL3 on oxidative stress and inflammatory response. Five groups (con, TNF‐α, TNF‐α + shNC, TNF‐α + shMETTL3, TNF‐α + shMETTL3 + oeRAC2) were divided in MH7A cells. A–D ROS A, SOD B, MDA C and T-AOC D were determined for evaluation of oxidative stress. E–G ELISA for level detection of IL-6 E, IL-1β F and IL-10 G. *P < 0.05, **P < 0.01, ***P < 0.001
Supplier Page from Abcam for Anti-RAC2 antibody