Supplementary Figure 8: Validation of METTL3 depletion in HEK293T cells.

(a) m⁶A immunoblotting of total RNA from wild type (WT) and METTL3-depleted cells (knockdown; KD) confirms loss of m⁶A. (b) Western blotting of WT and KD cells shows loss of METTL3 in KD cells. (c) IGV browser view showing stranded RNA-seq coverage at the METTL3 locus in METTL3 KD cells. Reads corresponding to METTL3 (antisense strand) are shown in purple. The expanded view shows the region in exon 3 of METTL3 which is targeted by the sgRNA being expressed in METTL3 KD cells. Yellow highlighted region indicates the site of one or two base insertions (T or TT) in the majority of reads. These insertions are predicted to cause a frameshift and premature stop codon. Results represent a minimum of n = 3 independent experiments.