Fig. 4: Hakai regulates m6A levels and m6A-dependent splicing events.
From: Hakai is required for stabilization of core components of the m6A mRNA methylation machinery
a LC–MS/MS quantification of m6A levels in either control samples or mRNA extracts depleted for the indicated proteins in S2R+ cells. The bar chart shows the mean with standard error (SE) of three biological replicates and three technical measurements. KD of Hakai results in substantial reduction of m6A levels. ∗∗∗P = 7.49E−04 (Hakai KD), ∗∗∗∗P = 7.51E−06 (Mettl3, Mettl14 KD). Unpaired two-tailed Student’s t-test for unequal variances. b Relative isoform quantification of m6A-regulated genes (fl(2)d and Hairless) upon depletion of the indicated components. The bar chart shows the mean with standard error (SE) of three biological replicates and three technical measurements. Hakai is required for m6A-dependent splicing regulation. c Number of differentially spliced events upon knockdown of the indicated proteins (left) and common differentially spliced targets (right) (FDR < 0.1). d Radar charts display relative distribution of differentially spliced events upon knockdown of Fl(2)d and Hakai. Alternative 5’splice site (A5SS) selection and intron retention (RI) are overrepresented events upon loss of m6A writer components. Source data for m6A measurement, qPCR and RNA-seq are provided as a Source Data file.
