Figure 6

GSDMD-N non-selectively increase plasma membrane permeability. (A-C) Intracellular ion concentration were monitored in GSDMD-N-HBD^*-HA-expressing CHO cells treated with 4-OHT. Sodium indicator (ANG-2, A), calcium indicator (Fluo-4, B) and potassium indicator (APG-2, C) were used together with PI. For each ion indicator, a series of representative time-lapse images were shown in the left panel and relative intensity change of ion indicator and PI fluorescence in one representative cell was graphed over time in the right panel. (D) GSDMD-N-HBD^*-HA-expressing CHO cells were treated with 4-OHT for 0.5 h in PBS (CTRL) or PBS containing 30 mM PEG400, PEG1450, PEG4000 or PEG8000. Cells were stained by PI and analyzed under microscope. (E) GSDMD-N-HBD^*-HA-expressing CHO cells were treated with 4-OHT in PBS or in PEG4000 PBS for 0.5 h, or were treated with 4-OHT in PEG4000 PBS for 0.5 h and then were changed into PBS for another 0.5 h. Cells were stained with PI and analyzed under microscope. Results shown are mean ± SD from two independent experiments. Scale bar, 10 μm.