Fig. 4: DVL2-mediated PCP signaling pathway is the target of METTL3 in HMC.

a Gene expressions in ARC and HMC (n = 10, WNT3 p = 0.8708, WNT11 p = 0.0046 and DVL2 p < 0.0001). b Protein expressions in ARC and HMC (n = 3, WNT11 p = 0.0464, DVL2 p = 0.0008). c The m6A modification levels in ARC and HMC (n = 3, WNT11 p = 0.0004, DVL2 p = 0.0034). d Upper: Western blot of co-precipitated proteins in endogenous METTL3 RIP analysis. Lower: qRT-PCR of co-precipitated mRNAs using METTL3 antibody in SRA 01/04 cell RIP analysis. IgG served as isotype control. (n = 3, WNT11 p > 0.9999, DVL2 p < 0.0001). e The immunofluorescent staining of WNT11 and DVL2 in ARC and HMC (n = 3, WNT11 p = 0.0187, DVL2 p = 0.0056). Yellow arrows indicate the enlarged view of DVL2’s polarized localization. Scale bar: 50 μm. f Protein expressions in ARC and HMC (n = 3, β-catenin p = 0.9594, p-JNK p = 0.0091). g Gene expressions in METTL3 OE cells (n = 4, WNT11 p = 0.9346, DVL2 p = 0.0121). NC = negative control. h Protein expressions in METTL3 OE cells (METTL3: n = 4, p = 0.0003; DVL2: n = 3, p = 0.0009; WNT11: n = 5, p = 0.5131). i p-JNK protein expressions in METTL3 OE cells (n = 3, p = 0.0281). j Protein expressions in cells transfected with siMETTL3 #1 (n = 3, METTL3 p = 0.0153, DVL2 p = 0.0084, p-JNK p = 0.0153). The m6A modification levels of DVL2 mRNA in METTL3 OE (k) or siMETTL3 (l) cells (n = 3, k: METTL3 OE p = 0.0001; l: siMETTL3 p < 0.0001). Data present are mean ± SD. The band density in Western blot was normalized to loading control as a ratio for statistical analysis. Level of significance was detected using two-sided Student’s t test (i, k, l) or two-way ANOVA with Šídák’s multiple comparisons test (a–h, j). n = biological replicates, ns. = not significant, ****p < 0.0001, ***p < 0.001, **p < 0.01 and *p < 0.05.