Figure 4

Deletion of calmodulin-binding sites on TRPV1 effects TRPV1 and AKAP150 association. (A) Co-immunoprecipitation and Western-blot analysis of AKAP150 with TRPV1, TRPV1ΔN CaM (TRPV1 mutant with deleted N-terminal calmodulin binding site) or TRPV1ΔC CaM (TRPV1 mutant with deleted C-terminal calmodulin binding site) in CHO cells. Results are representative of 4 independent trials. (B) Quantified densitometry of data represented in (A), n = 3. Significance with AKAP150 + TRPV1 transfection paradigm determined by determined by two-tailed, paired t-test, **p < 0.01. (C) Data from calcium-imaging traces of capsaicin (CAP)-stimulated (50 nM, 30 sec) intracellular Ca²⁺ accumulation from CHO cells expressing AKAP150 and TRPV1, TRPV1ΔN CaM TRPV1ΔC CaM are quantified as ΔF340/F380, mean ± SEM shown, n shown for each treatment/transfection paradigm. White asterisks indicate significance between AKAP150 and TRPV1ΔCCaM expressing cells and AKAP150 and TRPV1 expressing cells. (D) Data in C are transformed to illustrate percent desensitization. Significance determined by one-way ANOVA with Bonferroni correction, NS = no significance, ***p < 0.001.