Figure 3

Nebulin mutations affect binding to tropomyosin. Purified GST-nebulin domains bound to beads were incubated with purified α-tropomyosin and β-tropomyosin, then beads were washed and the bound proteins run in SDS-PAGE gels, and stained with Coomassie Blue. The relative intensity of bound α-tropomyosin and β-tropomyosin was quantified from three independent experiments. The mean value and standard deviations from three experiments are shown in the bar charts to the left, and gel pictures of representative experiments are shown on the right. Nebulin domains containing the p.Thr7382Pro (ex151m) mutation showed significantly weakened affinity for tropomyosin than wild-type (WT) proteins (P = 0.039). P values were calculated using the Kruskal–Wallis test when comparing three groups (S9) and the Mann–Whitney test when comparing two groups (S14, S18, S22). Asterisks indicate significant differences compared with the WT protein.