Figure 2.

Murepavadin induces intracellular Ca²⁺ mobilization, degranulation and chemokine production in MCs through MRGPRX2. (A) Calcium mobilization measurement following murepavadin (10 μM) stimulation of Fura-2 loaded WT-RBL and RBL-MRGPRX2 cells. (B) WT-RBL and RBL-MRGPRX2 cells were exposed to 10 μM murepavadin (30 min), and degranulation was assayed by measuring the release of β-hexosaminidase. (C) RBL-MRGPRX2 cells were incubated in the presence or absence of pertussis toxin (PTx), at a concentration of 100 ng/mL for 16 h and exposed to indicated concentrations of murepavadin (30 min) and degranulation was assayed by measuring the release of β-hexosaminidase. (D, E) WT-RBL and RBL-MRGPRX2 cells were incubated in the presence or absence of pertussis toxin (PTx) at a concentration of 100 ng/mL for 16 h and exposed to 10 μM of murepavadin (24 h) and the production of cytokine TNF-α and chemokine CCL2 were quantified by ELISA. Data presented are the mean ± SEM of at least three experiments. Statistical significance was determined by two-way ANOVA with Šídák’s or Tukey’s multiple comparisons at a value *p < 0.05, ***p < 0.001, ****p < 0.0001, and ns denotes “not significant”.