FIGURE 4.

SUCNR1 ligation promotes cytokine release from resting and IgE‐receptor activated mast cells. (A‐E) CBMC were pre‐incubated with succinate or cES, followed by 24 h incubation with 0.4 μg/ml anti‐IgE. Afterward, the supernatants were collected and analyzed by Bio‐plex assays (n = 8 and n = 4) and 17‐plex cytokine assay (n = 10). (F‐G) qPCR analysis of TNF and IL‐8 mRNA expression in IgE‐sensitized CBMC, pre‐incubated 30 min with or without 100 nM YM254890 or 1 μM cyclosporin A, followed by incubation of succinate (n = 4). (H) qPCR analysis of TNF mRNA in control and SUCNR1 KD LAD‐2 cells pre‐incubated with vehicle succinate or cES, followed by 24 h incubation of 100 μg/ml anti‐IgE (n = 4, normalized to β‐actin expression). (I‐J) Multiplex cytokine analysis of CBMC incubated with 1 µM XT1, followed by succinate and anti‐IgE (0.4 μg/ml, 24 h) (n = 8). Paired student's t‐test (C, I‐J), RM one‐way ANOVA with Dunnett's multiple comparison test (F‐G), and RM two‐way ANOVA with Sidak's multiple comparison test (H) were applied. *p < 0.05, **p < 0.01. ***p < 0.001. ****p < 0.0001