Fig. 6

Ac2-26 enhances interaction between CaM and TRPV1 via FPR2. a Representative images of double immunofluorescent staining after application with scramble (3.3 μM), Ac2-26 (3.3 μM) and Boc2 (10 μM) + Ac2-26 (3.3 μM) in cultured AnxA1^−/− DRG neurons. The areas in the dotted box in merge pictures zoom in and present in magnify panels. Scale bar, 50 μm in TRPV1, CaM and merge pictures, 25 μm in magnify pictures. b Quantification of the fluorescent intensities of TRPV1 positive and CaM positive (TRPV1⁺/CaM⁺) cells among scramble, Ac2-26 and Boc2 + Ac2-26 groups. (Ac2-26 versus scramble group, ****P < 0.0001; Ac2-26 versus Boc2 + Ac2-26 group, ***P < 0.001, one-way ANOVA, post hoc Student’s t test. n = 10 in each group). All data are represented as mean ± SD. c and d Representative Western blots (WB) band images of immunoprecipitation (IP) experiments in cultured AnxA1^−/− DRG neurons treated with scramble (3.3 μM), Ac2-26 (3.3 μM) and Boc2 (10 μM) + Ac2-26 (3.3 μM), respectively. c TRPV1 antibody incubated immunoprecipitations were detected with CaM and TRPV1 antibodies by western blotting. d CaM antibody incubated immunoprecipitants were detected with TRPV1 and CaM antibodies by western blotting. Input, 10 μg protein of the extracts without IP was loaded. IgG, immunoprecipitants incubated with nonspecific IgG (IgG) as negative control. e Representative traces of capsaicin-evoked currents with co-application of scramble, Ac2-26 and Boc2 + Ac2-26 in cultured AnxA1^−/− DRG neurons. f Quantification of the fold change peak current in scramble, Ac2-26 and Boc2 + Ac2-26 groups. The data of fold change peak current in scramble group were normalized to 1 (Ac2-26 versus scramble group, ****P < 0.0001; Ac2-26 versus Boc2 + Ac2-26 group, ****P < 0.0001, one-way ANOVA, post hoc Tukey’s multiple comparisons test, n = 8 in each group)