Fig 2: IKCs signaling is impaired in the R312H brain.A Representative Western blots of major components of IKC signaling (total and phosphorylated, or in the case of Ras, GTP-bound) from the brains of the indicated genotypes. Loading control: Bradford assay. B Representative co-IPs of KCNB1 (IP) with the indicated adhesome proteins. Samples were loaded to have approximately the same amount of KCNB1 protein per genotype. The same membrane was used to visualize Vinculin and Talin-1 (membrane stripping). C Representative co-IPs of KCNB1 (IP) with the indicated phosphorylated adhesome proteins: Paxillin phosphorylated at Tyr118 (pPax), Talin-1 phosphorylated at Ser425 (pTal), and Vinculin phosphorylated at Tyr100 (pVin). Samples were loaded to have approximately the same amount of KCNB1 protein per genotype. The same membrane was used to visualize pVinculin and pTalin-1 (membrane stripping). Controls: mouse IgG and empty beads. D Densitometric quantifications of 3–4 experiments as those shown in (C). *P < 0.05, **P < 0.01, and ***P < 0.001 (one-way ANOVA, Tukey’s post hoc).