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. 2020 Apr 24;9(4):1061. doi: 10.3390/cells9041061

Figure 1.

Figure 1

Characterization of RNA methyltransferase Mettl3 interactions with 5′-ETS of 47S pre-rRNA. (A) Scheme of the RT-qPCR primer locations on the pre-47S rRNA. (B) Relative amounts of rRNA fragments in immunoprecipitates obtained with anti-m6A antibodies from control and Mettl3 KD cells, quantified by RT-qPCR analysis (* p < 0.05, ** p < 0.01, *** p < 0.001). Negative control (unmethylated RNA from the EpiMark® N6-Methyladenosine Enrichment Kit) A’-A0 (primers for amplification of A’-A0 spacer), 5′-ETS (primers for amplification of the part of 47S pre-rRNA before A’), nucleolin (m6A methylated mRNA based on MeT-DB2.0 browser data). (C) Enrichment of rRNA fragments in immunoprecipitates obtained with anti-Mettl3 antibodies from control cells, quantified by RT-qPCR analysis. (D) Scheme of 5′-ETS in rRNA with designated adenosines in the context of predicted DRACH sequences. (E) PCR analysis of T4 RNA ligase assay for pre-rRNA from control and Mettl3 KD cells (visualization of fragments separated using gel-electrophoresis and their relative quantification).