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. 2014 May 16;307(2):L173–L185. doi: 10.1152/ajplung.00083.2014

Fig. 6.

Fig. 6.

Autophagy mediates NLRP3 inflammasome activation during mechanical ventilation. BMDMs (MΦ) isolated from donor mice were transfected with a scrambled siRNA (siSc) or ATG5 siRNA (siAtg5). After 48 h, the efficiency of transfection was evaluated by Western blot. Mouse AMs were depleted with CLOD as described in materials and methods. BMDMs transfected with a siSc or siAtg5 were intravenously injected into AM-depleted mice. Mice were ventilated with a high tidal volume (MV, 28 ml/kg) for 2 h. At the end of experiments, lung tissues were recovered. A: effects of mechanical ventilation on NLRP3 inflammasome activation in the lung. The assembly of NLRP3 inflammasome was detected using immunoprecipitation (IP) with anti-NLRP3 antibody followed by immunoblotting (IB) for ASC, NLRP3, and caspase-1 cleavage product p10 fragments (Casp-1). B: relative densities of the bands of ASC proteins. Bar graph shows the relative abundance of ASC protein (normalized to that of NLRP3) from 3 experiments. C: relative density of the bands of Casp-1. Bar graph shows the relative abundance of Casp-1 protein (normalized to that of NLRP3) from 3 experiments. *P < 0.05 vs. the control (nonventilated and nontreated) group, †P < 0.05 vs. MV alone group, ‡P < 0.05 compared with siSc group.