Figure 2

Mutations affect the binding of nebulin to F-actin. Nebulin protein domains were incubated with F-actin and centrifuged. Pellet and supernatant fractions were separated, run on SDS-PAGE gels, and stained with Coomassie Blue. The relative abundance of nebulin protein in the pellet 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 mutation p.Ser6366Ile (ex122m) showed significantly strengthened actin affinity (P = 0.048). 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 wild-type (WT) protein.