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. 2020 Apr 24;11:703. doi: 10.3389/fimmu.2020.00703

FIGURE 1.

FIGURE 1

Osthole inhibits MRGPRX2-induced Ca2+ mobilization and degranulation in LAD2 mast cells. (A,C,E) Intracellular Ca2+ mobilization and (B,D,F) degranulation in LAD2 human mast cells was determined following pre-incubation with vehicle (0.2% DMSO in PBS) or varying concentrations of osthole for 30 min. Cells were stimulated with the MRGPRX2 agonist (A) compound 48/80 (100 ng/ml), (C) substance P (300 nM) or (E) LL-37 (3 μM) and changes in fluorescence intensities were recorded for 90 s. Data are plotted as the change in fluorescence intensity [minimum (Min) subtracted from maximum (Max) value] measurements. Vehicle- or osthole-treated cells were exposed to (B) compound 48/80, (D) substance P or (E) LL-37 and degranulation was quantified by β-hexosaminidase release. Values are plotted as percentages of total cell lysate β-hexosaminidase content. Data shown are mean ± SE of three independent experiments. Statistical significance was determined by unpaired Student’s t-test with values compared between the osthole- and vehicle-treated groups *p < 0.05 and **p < 0.01.