Fig. 4

Anantin facilitates P2X3 localization to lipid rafts. a Top panel is a representative example of Western immunoblotting showing the amount of P2X3 receptors in raft (R) and non-raft (NR) membrane fractions in control and after treatment with anantin (500 nM, 24 h), MβCD (10 mM, 30 min) or their combination; flotillin bands indicate lipid raft membrane fractions. Lower panel shows total amount of P2X3 receptors for each experimental condition; β-tubulin was used as loading control of the total extract. Histograms at the bottom quantify mean P2X3 relative optical density values in lipid raft and non-raft membrane fractions (n = 4; *p < 0.05, Kruskal–Wallis test). Anantin relocates P2X3 receptors from non-raft to lipid raft membrane compartments, while MβCD abolishes anantin effect. b Top panel is a representative example of Western immunoblotting showing the amount of P2X3 receptors in raft and non-raft membrane fractions in control and after treatment with anantin (500 nM, 2–3 h), siRNA BNP (48 h), or their combination; flotillin bands indicate raft membrane fractions. Lower panel shows total amount of P2X3 receptors in each experimental condition; β-tubulin was used as loading control of the total extract. Histograms at the bottom quantify P2X3 relative optical density in lipid raft and non-raft membrane fractions according to the used treatment (n = 4; *p < 0.05, Kruskal–Wallis test). c Histograms show mean P2X3 current amplitude values in control and after application of MβCD (10 mM, 30 min), anantin (500 nM, 2–3 h), or their combination (n = 47, 58, 40, 61; *p < 0.05, Kruskal–Wallis test). Representative current traces of P2X3 receptors evoked by α,β-meATP (10 µM, 2 s) for each experimental condition are shown on the right. MβCD per se decreases P2X3 current density, whereas MβCD co-application with anantin returns current density values to control level