CASK/P2X3 complex in KI trigeminal neurons. A, Example of immunopurified P2X3 from trigeminal ganglia probed with anti-CASK antibodies reveals more abundant CASK/P2X3 complex in KI ganglion cultures than in WT ones. Histograms quantify this effect (n = 5, p = 0.038). Ab, indicate signal from unrelated antibody. B, CASK/P2X3 co-immunoprecipitation experiments from ganglion cultures demonstrate that CASK/P2X3 complex is sensitive to ω-Agatoxin IVA treatment (400 nM, overnight; Aga). Note the more intense CASK/P2X3 signal in KI samples as seen in A. P2X3 immunoprecipitation input and β-Tubulin expression shown as gel loading controls. C, CASK/P2X3 complex is sensitive to KN-93 pre-incubation (5 μM, 90 min). D, Examples of anti-phospho-CaMKII T286 Western immunoblots of extracts from WT and KI trigeminal cultures in control conditions and after CASK silencing. β-Tubulin is shown as loading control. Histograms quantify the effect (#, p = 0.05; *p = 0.03 for WT and p = 0.025 for KI; n = 3).