Fig. 4

Injurious mechanical ventilation activated the pyroptosis effector GSDMD. A, B Western blot of the neonatal rat brain for caspase-1 (A) and GSDMD (B) showing increased relative band intensity of the active GSDMD fragment in whole brain homogenates from animals ventilated with high Vt. N = 4 animals/group (caspase-1) and N = 10 animals/group (GSDMD). Data was analyzed by ANOVA with Dunnett’s post-test, p values < 0.05 are shown on bars. Data are presented as box plots showing the median, 25th to 75th percentile, and minimal to maximal value normalized to control. C–E Photomicrographs (20×; bars = 50 µm) of representative brain sections immunostained for the GSDMD showing the corpus callosum and subcortical white matter in controls (C), low Vt (D), and high Vt (E) animals. There was increased staining for GSDMD in the corpus callosum and subcortical white matter of animals ventilated with high Vt. N = 4 animals/group. Data was analyzed by ANOVA with Dunnett’s post-test for multiple comparison, p values < 0.05 are shown on bars