Figure 4.

Effects of atg5 deficiency on LC3 expression and autophagy flux H₂O₂-activated atg5^−/− platelets of mice. (A) Schematic for platelet-specific atg5^−/− mice generation by using Cre-loxP recombination system. (B and C) Identification of platelet-specific atg5^−/− mice by PCR (B) and western blotting (C). (D) Phenotype of the representative 8-week-old mice of WT, Atg5^f/f, and platelet-specific atg5^−/− mice. (E) Hematological parameters of WT, Atg5^f/f, and platelet-specific atg5^−/− mice analyzed using IDEXX Procyte Dx. Values are expressed as the means ± S.E.M. (n = 7). (F) Washed mouse Atg5^f/f and atg5^−/− platelets (1 × 10⁹/mL each) treated with or without 100 μM H₂O₂ for 60 min, and LC3 was then detected by western blotting. (G) Endogenous LC3 after 100 μM H₂O₂ treatment detected through immunofluorescence (IF) under a deconvolution microscope. The arrows indicate LC3 puncta. Scale bar: 1 μm. The profiles (C and G) are representative examples of 4 similar experiments. Data (F) are presented as the means ± S.E.M. (n = 4)