Fig. 2

Identification of METTL3 targets via MeRIP-seq and RNA-seq. a, Immunoblotting of METTL3 in SW480 and SW620 cells (left), and in METTL3 knockdown SW620 and control SW620 cells (right). b, Distribution of peaks (fold change > 1.5 or < − 1.5, P < 0.05) with a significant change in both the RNA expression level and m⁶A level in SW620 cells compared with SW480 cells (left), and in METTL3 knockdown SW620 cells compared to control SW620 cells (right). c, Venn diagram showing the shared peaks between metastatic-related hyper-up peaks and METTL3-related hypo-down peaks. A total of 192 shared peaks corresponding to 158 specific genes were observed. d, GO biological process enrichment analysis of the above shared peaks. e, The m⁶A abundances in SEMA3A, BCHE, ZFP36L2, and SOX2 transcripts in SW620 cells related to the SW480 cells (left panel), and in METTL3-knockdown SW620 cells (shMETTL3#1) related to the control SW620 cells (shNC) (right panel). f, Gene-specific m⁶A qPCR analysis of alterations in the m⁶A level in four representative genes in SW620 and SW480 cells. g, Gene-specific m⁶A qPCR analysis of alterations in the m⁶A level in four representative genes in METTL3-knockdown SW620 and control SW620 cells. h, Immunoblotting assay of SOX2 after METTL3-knockdown in SW620 and HCT116 cells. The data in f, and g are presented as the mean ± SDs (n = 3). *P < 0.05, **P < 0.01 (Student’s t-test). β-Actin was used as the loading control. The relative m⁶A level was normalized by input. The relative expression level was normalized by the β-Actin