Figure 5
From: Dissecting domains necessary for activation and repression of splicing by muscleblind-like protein 1
MBNL binding to Vldlr and Tpm1 RNAs. RNA binding was assessed by native gel electrophoretic mobility shift assay. RNAs are the MBNL-responsive element from Vldlr (left panels), the upstream MBNL binding site of Tpm1 exon 3 (middle panels), and the downstream MBNL binding site of Tpm1 exon 3 (right panels). Recombinant proteins used were: A, MBNL-N, the wild-type MBNL construct comprising amino acids 2–253 and containing all four zinc fingers, B, MBNL-N-M34, with ZF34 mutated, C, MBNL-N-M12 with ZF12 mutated, D, MBNL-2-116 E, MBNL-2-91 F, MBNL-2-72. Increasing protein concentrations are indicated by the wedges above each panel. Protein concentrations were 0, 0.1, 0.5, 1, 2, 5, 10, 25, 50, 100, 250, 500 nM for panel A, 0, 0.1, 0.5, 1, 2, 5, 10, 25, 50, 100, 250, 1000 nM for panels D and E, and 0, 1, 2, 5, 10, 25, 50, 100, 250, 500, 1000, 2000 nM for panels B, C and F. Estimated Kd’s are indicated in the lower right corner of each panel.