Increased accumulation and release of proapoptotic factors in GSD-Ib neutrophils. Peritoneal neutrophils were isolated from 6- to 7-week-old unaffected (+/+) and GSD-Ib (−/−) mice during thioglycollate-elicited peritonitis. (A) Representative immunofluorescence of Smac/Diablo staining (green fluorescence), Hoechst 33342 nuclei staining (blue fluorescence), and MitoTracker Red mitochondrial staining (red fluorescence), magnification, ×1000, and quantification of Smac/Diablo-positive neutrophils in unaffected and GSD-Ib mice. (B) Western-blot analysis of protein extracts of neutrophils using antibodies against Smac/Diablo, β-actin or GAPDH. Each lane contains 50 μg protein. (C) Representative immunofluorescence of Omi/HtrA2 staining (green fluorescence), Hoechst 33 342 nuclei staining (blue fluorescence), and MitoTracker Red mitochondrial staining (red fluorescence), magnification, ×1000, and quantification of Omi/HtrA2-positive neutrophils in unaffected and GSD-Ib mice. Data represent the means (± SEM) of 3 independent experiments. ***P < .001. (D) Western blot analysis of protein extracts of neutrophils using antibodies against Omi/HtrA2, β-actin, or GAPDH. Each lane contains 50 μg protein.