Extended Data Fig. 4: METTL3 chromatin binding is dependent on its own catalytic activity.
From: METTL3 regulates heterochromatin in mouse embryonic stem cells
a, ChIP–qPCR showing binding patterns of METTL3 on IAPEz-int elements in parental, Mettl3 KO and rescued cell lines with METTL3WT or METTL3APPA. The mean of three biological replicates ± s.d. is shown. *P < 0.05, **P < 0.01, two-sided t-test. Exact P values are provided in the Source Data. b, Aggregation plots showing METTL3 enrichment levels on IAPEz-int in parental, Mettl3 KO and rescued cell lines with METTL3WT or METTL3APPA. c, Western blot showing interactions of METTL14 with reintroduced METTL3 (METTL3WT or METTL3APPA) in Mettl3 KO cells. d, Aggregation plots showing METTL3 enrichment levels on IAPEz-int in Mettl3 KO rescued cells with METTL3WT, METTL3W475A or METTL3N477A. e, Western blot showing METTL3 protein levels in parental, Mettl3 KO and rescued cell lines with METTL3WT, METTL3APPA, METTL3W475A or METTL3N477A. f, Aggregation plots showing METTL3 enrichment levels on IAPEz-int in parental and Mettl14 KO cell lines. g, Western blots showing METTL14 protein levels in parental and Mettl14 KO cell lines. h, Aggregation plots showing METTL3 enrichment levels on IAPEz-int in parental and Rbm15/15b DKO cell lines. i, Western blots showing RBM15 and RBM15B protein levels in parental and Rbm15/15b DKO cell lines. j, Aggregation plots showing METTL3 enrichment levels on IAPEz-int in control and m6A methyltransferase complex components KD cell lines. k, Western blots showing protein levels of m6A methyltransferase complex components in control and KD cell lines. l, A cartoon illustrating the dCas9–METTL3 tethering assay in Mettl3 KO cell lines. m, Western blot showing Cas9 and METTL3 protein levels upon Dox treatment. n, ChIP–qPCR of Cas9 (left) and H3K9me3 (right) on IAPEz and control regions. The mean of three biological replicates ± s.d. is shown. *P < 0.05, **P < 0.01, two-sided t-test. Exact P values are provided in the Source Data. Uniquely + randomly mapped ChIP–seq reads were used in b, d, f, h, j. Blots are representative of two independent experiments in c, e, g, i, k, m. For blots source data, see Supplementary Fig. 1. MTA and MaSat used in ChIP–qPCR are examples of repetitive elements not bound by METTL3.
