FIGURE 2.

MRGPR-X1-induced TRPV1 activation is independent from PKC activity (manganese influx). Mn²⁺-induced quenching of fura2 fluorescence in F11-MRGPR-X1 (A) or F11-MRGPR-X1/TRPV1 cells (B and C) were measured. BAM8–22 (2 μm, final) or HBS (basal) were manually injected at time point 25 s. In A and B cells were preincubated or not with BCTC (500 nm, 5 min), and in C cells were preincubated either with the carrier DMSO (0.1%, 30 min) or with BIM-X (1 μm, 30 min). Data from 3–5 independent experiments were compiled and normalized by defining the first value (0.25 s) measured as 100% and expressed as the mean ± S.E. In A and B, differences in Mn²⁺-induced quenching of fura2 fluorescence before application and at time point 45 s are also shown in the bar graphs. In C, differences between basal (DMSO or BIM-X) and BAM8–22-provoked Mn²⁺-induced quenching of fura2 fluorescence at time point 45 s are given. Asterisks indicate a significant difference (***, p < 0.001) between BAM8–22-stimulated and unstimulated cells; hash signs (###, p < 0.001) between BCTC-treated and untreated cells.