Figure 4.

The inhibition of TRPM2 channel reduced PL-induced Ca²⁺ entry. (A) PL-induced Ca²⁺ entry was dramatically reduced in the presence of 10 µM Econazole (30 min preincubation). PL was added at 20% v/v. Data are means ± s.e.m. of [Ca²⁺]_i traces recorded in different cells. Number of cells: PL alone: 40 cells from 3 experiments; PL + Econazole 10 µM: 50 cells from 3 experiments. (B) Mean ± s.e.m. of the Ca²⁺ response to 20% v/v PL recorded at the peak and at the plateau under the designated treatments. Data are means ± s.e.m. of [Ca²⁺]_i measured by confocal imaging at peak maxima. Number of cells: PL alone: 40 cells from 3 experiments; PL + Econazole 10 µM: 50 cells from 3 experiments. Asterisks on bars indicate statistical differences determined by two-way ANOVA with Bonferroni’s correction (p < 0.001). (C) Expression of TRPM2 gene in bEND5 cells after RNAi. The mRNA quantity of TRPM2 was determined by qRT-PCR and is represented as mean relative expression ± SD (n = 3, * p < 0.001, t-test). (D) PL-induced Ca²⁺ entry was completely abrogated in cells transfected with RNAi targeting TRPM2. PL was added at 20% v/v. Data are means ± s.e.m. of [Ca²⁺]_i traces recorded in different cells. Number of cells: PL alone: 30 cells from 3 experiments; PL after RNAi for TRPM2: 50 cells from 3 experiments. (E) Mean ± S.E.M. of the Ca²⁺ response to 20% v/v PL recorded at the peak and at the plateau under the designated treatments. Data are means ± s.e.m. of [Ca²⁺]_i measured by confocal imaging at peak maxima. Number of cells: PL alone: 30 cells from 3 experiments; PL after RNAi for TRPM2: 50 cells from 3 experiments. Asterisks on bars indicate statistical differences determined by two-way ANOVA with Bonferroni’s correction (p < 0.001).