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. 2020 Apr 3;11:229. doi: 10.3389/fimmu.2020.00229

Figure 6.

Figure 6

Hepatocytes release high-mobility group box-1 (HMGB1) in exosomes. Immunoblots for CD81, TSG101, and HMGB1 in exosomes isolated from (A) cell culture media from wild-type (WT) hepatocytes treated with lipopolysaccharide (LPS) (1 μg/ml) for 24 h and (B) plasma from WT mice intraperitoneally injected with LPS (5 mg/kg) for 4 h. NanoSight™ analysis of exosomes isolated from (C) cell culture media and (D) mouse plasma. (E,F) Immunoblots for HMGB1 in the supernatant and cell lysates of hepatocytes treated with GW4869 or spiroepoxide for 2 h, then challenged with LPS (1 μg/ml) for 24 h. (G) Plasma HMGB1 level in WT mice injected intravenously with GW4869 (2.5 mg/kg) for 1 h prior to intraperitoneal injection of LPS (5 mg/kg) for 4 h. (H) Plasma HMGB1 level in WT mice pretreated for 48 h with scrambled (control) or Rab27a-targeted shRNA via intravenous injection followed by intraperitoneal injection with LPS (5 mg/kg) for 4 h. Each point represents one mouse. *P < 0.05. n = 3.