Figure 3. rs20544 C/T polymorphism affects FMRP binding to MMP‐9 RNA molecule.

1. Representative RNA electrophoretic mobility shift assay (REMSA) results. Labeled MMP‐9 RNA probe was incubated in the absence (lanes 1 and 8) or presence of increasing amounts of purified FMRP (lanes 2–6 and 9–13). 20× molar excess of unlabeled probe was added as a competitor to confirm the specificity of the interaction (lanes 7 and 14).
2. Scheme of MMP‐9 mRNA indicating the location of the rs20544 polymorphism and part of the sequence that was used as a probe in the REMSA (A, C) and filter binding assay (D).
3. Quantification of REMSA experiments. The relative mobility change of the protein–RNA complexes from the corresponding free probe band was plotted against increasing FMRP concentrations. For each FMRP concentration, the average distance of the shifted complex/free probe band was calculated from at least three independent experiments.
4. Quantification of filter binding assay. The fraction of bound RNA was plotted against increasing FMRP concentrations. The data are from five independent experiments. Each column represents the mean counted from range of concentrations (indicated in Materials and Methods), with the final concentration shown on the abscissa.

Data information: Error bars indicate the SEM. *P < 0.05, **P < 0.01 (Student's t‐test).