Fig. 3 | Laboratory Investigation

Fig. 3

From: NLRP3/ASC-mediated alveolar macrophage pyroptosis enhances HMGB1 secretion in acute lung injury induced by cardiopulmonary bypass

Fig. 3

Inhibiting AM pyroptosis by ASC silencing reduces the OGD-enhanced HMGB1 secretion in AM. Following transfection with ASC-specific siRNA or scramble (scrbl) siRNA, the cells were stained with Cy3-anti-HMGB1 and DAPI. The distribution of HMGB1 in the different groups of AM was characterized by confocal microscopy. In addition, the cytosolic and total proteins were extracted, and the relative levels of cytoplasmic and total HMGB1 in AM were determined by Western blot. The levels of HMGB1 in the cell supernatants were determined by ELISA. Data are representative images or expressed as the mean ± SEM of each group (n = 3 per group) from three separate experiments. a Immunofluorescent characterization of HMGB1 distribution in AM (magnification ×740). b Western blot analysis of the levels of cytoplasmic and total HMGB1 expression in AM. c The levels of HMGB1 in the supernatants. **p < 0.01 vs. the control group; ##p < 0.01 or #p < 0.05 vs. the OGD + ASC siRNA group

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