Extended Data Fig. 3. E-selectin triggers rapid S100A8/A9 release.
a Isolated neutrophils were treated for 10 min with PBS, E-selectin or E-selectin in the presence of high [K+]ex and the amount of full length GSDMD, GSDMD-NT and GAPDH was determined in the cell lysates (PBS control group and E-selectin stimulated group values and representative blot are the same as in Fig. 2g; n = 4 (control, E-selectin), 3 (E-selectin/ high [K+]ex) independent experiments). b Overall intracellular S100A8/A9 content was assessed by Western blotting in bone marrow neutrophils of WT and Kcna3-/- mice (n = 4 mice/group). c Isolated human neutrophils were stimulated with PBS (control) or E-selectin (1 µg ml-1) for 1 min, 5 min or 10 min and S100A8/A9 levels in the supernatants were assessed by ELISA (n = 3 (control 10 min, E-selectin 10 min), 4 (E-selectin 1 min), 5 (control 1 min, control 5 min, E-selectin 5 min) individual experiments. Values from the 10 min time point (control and E-selectin) are the same as in Fig. 3g. Data are presented as mean ± s.e.m. (one-way ANOVA, Tukey’s comparison for a, two-tailed unpaired student’s t-test for b and two-way ANOVA, Sidak’s multiple comparison for c), as representative Western blot for a, ns: not significant.