Fig. 5.

Evobrutinib specifically inhibits B cellular excitatory calcium mobilization and cytokine production. a, b. B cells were isolated from naïve C57/BL6 mice after 3 days of evobrutinib treatment and stained using Fluo-3 and Fura Red for calcium mobilization assay. After 25 s baseline recording, cells were stimulated with 20 µg/ml anti-IgM/anti-IgG. c Calcium mobilization on naïve murine B cells was analyzed after at least 30 min In vitro pre-incubation with the indicated concentrations evobrutinib. After 25 s baseline recording, cells were stimulated with the indicated concentrations anti-IgM/anti-IgG. d For BTKi reversibility, murine B cells were incubated for 30 min with the indicated concentrations evobrutinib and stained for calcium mobilization directly after as well as after 24 and 48 h. e Cytokine production was analyzed after 3 h anti-IgM/anti-IgG stimulation via qPCR. f B and T cells were pre-incubated with the indicated concentrations of evobrutinib and stimulated with anti-IgM/anti-IgG (B cells) or anti-CD3/anti-CD28 crosslinking (T cells). Median, n = 4–10, pooled from at least 2 independent experiments, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001