Fig. 2: METTL16 binds and methylates MAT2A to mediate MAT2A splicing and expression.
A Schematic illustration of METTL16 regulation of MAT2A expression in iSLK-RGB-BAC16 cells. B iSLK-RGB-BAC16 cells were transfected with siRNAs targeting METTL16 (siM16) or the control siRNA (siCtrl), and collected for RNA purification. The level of m6A on a specific transcript locus was examined by MeRIP-qPCR. m6A peaks located at the MAT2A hp1 site and hp2-6 sites were detected. Dicer m6A peak was used as a negative control. C iSLK-RGB-BAC16 cells were transfected with siRNAs targeting METTL16 or the control siRNA and collected for RNA purification. The expression levels of mature MAT2A and retained intron transcripts were examined by qRT-PCR. GAPDH was used as a normalization control. D iSLK-RGB-BAC16 cells were transfected with siRNAs targeting METTL16 or the control siRNA. Cells were lysed and the expression of MAT2A protein was detected by Western-blotting. GAPDH was used as a loading control. E iSLK-RGB-BAC16 cells transfected with a plasmid expressing METTL16 or a vector control were collected for RNA purification. The level of m6A on a specific transcript locus was examined by MeRIP-qPCR. m6A peaks located at the MAT2A hp1 site and hp2-6 sites were detected. Dicer m6A peak was used as a negative control. F The expression level of mature MAT2A in METTL16 overexpressed iSLK-RGB-BAC16 cells were detected by RT-qPCR. G iSLK-RGB-BAC16 cells transfected with a plasmid expressing METTL16 or a vector control were collected for RNA purification. RNA immunoprecipitation (RIP)-qPCR was performed and fold of enrichment of MAT2A RNA was calculated. *, **, and *** indicate p values of <0.05, <0.01, and <0.001, respectively, ns not significant.
