Fig. 8

IKK controls platelet-derived exosome secretion during septic shock. a–c Mice were treated with either 3% Tween 80 (5 mL/kg, vehicle) or 10 mg/kg BMS-345541 in 3% Tween 80 by oral gavage. Two hours post-treatment, sham or CLP procedures were conducted. n = 5 mice per group. a Exosomes were isolated from equal plasma volumes (100 μL), and the protein concentration was determined using a BCA protein assay kit. b Quantification of dsDNA in the plasma of mice using PicoGreen fluorescent dye. c Lung histology was assessed with H&E staining (magnification × 400). The sham group showed normal lung tissue with thin alveolar walls and few alveolar macrophages. Red arrows indicate neutrophils in the alveolar space, green arrows indicate neutrophils in the interstitial space, black arrows indicate alveolar macrophages, yellow arrows indicate hyaline membranes, and blue arrows indicate thickening of the alveolar walls. Lung injury scores were assessed. Scale bar, 50 μm. d–f Platelets were pre-incubated with IKK inhibitors (10 μM BMS-345541 or 25 μM BAY 11–7082) for 5 min and stimulated with LPS (1 μg/mL) for 2 h. n = 3. d The expression levels of p-IκB and IκB in platelets were measured by western blot. e Exosomal protein levels in the supernatant of stimulated platelets were measured using BCA. f HMGB1 expression in platelets was measured by western blot. Graphs represent means ± SEM; *P < 0.05, **P < 0.01 compared within two groups