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. 2024 Apr 20;38(6):1236–1245. doi: 10.1038/s41375-024-02222-w

Fig. 2. 8CA/8AA increase intron retention in MAT2A RNA by decreasing METTL16 occupancy.

Fig. 2

A Diagram of primers covering different regions of MAT2A RNA. B, C The ratios of intron retention in MV4-11 and KG-1a cells were measured after 24 h treatment with vehicle (NT) or 1 μM 8CA/8AA (n = 3). D–G The ratios of intron retention in MV4-11 and KG-1a cells were measured after 24 h pre-treatment with complete media or methionine-depleted media and followed by 24 h treatment with vehicle (NT) or 1 μM 8CA/8AA (n = 3). The respective protein levels of MAT2A were measured as well. H, I The binding of hp1 and hp2–6 clusters of MAT2A RNA and GAPDH RNA to METTL16 protein in MV4-11 and KG-1a cells were measured after 48 h treatment with vehicle (NT) or 1 μM 8CA/8AA (n = 3). J–M Protein and RNA levels of METTL16 in MV4-11 and KG-1a cells were measured after 48 h treatment with vehicle (NT) or 1 μM 8CA/8AA (n = 3). N, O MV4-11 and KG-1a cells were pretreated with vehicle (NT) or 1 μM 8CA/8AA for 24 h, and then incubated with 10 μg/mL puromycin for 10 min. Puromycin incorporation during nascent protein synthesis was measured through western blotting. P The components of METTL16 RNA in MV4-11 cells after 24 h treatment with vehicle (NT) or 1 μM 8CA/8AA in different fractions extracted from the polysome profiling assay were measured by qRT-PCR and normalized to inputs (n = 3). nsp > 0.05, *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, ****p ≤ 0.0001.