Agonist-induced LC3II reduction involves autolysosomal proteolysis. (A) Western blot analysis of LC3II changes in response to agonists in banked human platelets pretreated with a PIC. Platelets were incubated with the PIC for 60 minutes before 30-minute agonist stimulation with thrombin (0.1 U/mL), PAR1 peptide (100 µM), and CVX (0.1 µg/mL), respectively. The images shown are representative of at least 3 independent experiments. (B) Western blot analysis of LC3II in thrombin-stimulated (0.1 U/mL, 30 minutes) mouse platelets after preincubation with NH4Cl (20 mM, 2 hours). The images shown are representative of 4 independent experiments. *P < .05. (C) Western blot analysis of LC3II in thrombin-stimulated (0.1 U/mL, 30 minutes) platelets isolated from mice lacking VAMP2, VAMP3, and/or VAMP8, and from wild-type control mice. The images shown are representative of 2 independent experiments.
