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. 2020 Jun 4;48(13):7421–7438. doi: 10.1093/nar/gkaa475

Figure 1.

Figure 1.

DBHS proteins are preferentially enriched by GGGGCC repeat RNA in the G-quadruplex conformation. (A) Comparative enrichment of DBHS-family proteins and hnRNP K by (GGGGCC)4 RNA and (CCCCGG)4 RNA. SILAC was used for comparative analysis of proteins enriched by pulldown with biotinylated RNA oligonucleotides from HEK293T lysate (62). Enrichment by G-rich ‘Sense’ relative to enrichment by C-rich ‘Antisense’, as indicated by SILAC ratios of (GGGGCC)4 / (CCCCGG)4, is plotted; a log2 scale is used to illustrate strand preference. (B) Comparative enrichment of DBHS-family proteins by (GGGGCC)4 RNA in G-quadruplex and non-G-quadruplex conformations, shown relative to enrichment by (CCCCGG)4 RNA. SILAC ratios for (GGGGCC)4/(CCCCGG)4 are shown for both (GGGGCC)4 conformations. (C) Western blot validation of RNA pulldown results. 2.5% input volume was loaded for reference (‘In’). Pulldown with bead-only control (‘B’) was compared to antisense (‘AS’; (CCCCGG)4) RNA and ‘Sense’ ((GGGGCC)4) RNA in non-G-quadruplex and G-quadruplex conformations. Results shown are representative of three replicates.