MDH2 predominantly binds cytosolic RNAs. (A) Sunburst chart showing the distribution of cross-linking counts in binding sites identified from MDH2 eCLIP data. “Cytosolic” indicates nuclear-encoded RNAs and “Mito” indicates mitochondrially transcribed RNAs. (B) Cross-linking (CL) count distribution of selected target RNAs bound by MDH2 visualized using the IGV interactive tool (Robinson et al. 2011). (C) Representative electromobility shift assay (EMSA) assays were performed to assess the affinity of MDH2 to a tRNA Arg TCT 3-2 probe or a control RNA probe (RPS23 5′-UTR sequence). Binding of eGFP to the target RNA probe is included as a negative control. (D) Binding affinities from EMSA experiments were estimated using a nonlinear regression fit (solid line). All shifted bands (all bands above the free migrating RNA) were included in the quantification of total RNA binding. Data are reported as mean ± SD (dots with error bars). Number of experiments, n = 3–4 from two independent protein purifications.
