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After that, the membranes including the goal proteins were incubated with the corresponding antibody including anti-ANP antibody (abcam, Cambridge, MA, USA) diluted 1: 5000, anti-MYH7 antibody (Proteintech Group, Inc, Wuhan, China) diluted 1: 2000, anti-COXII antibody (Cell Signaling Technology, Inc, Boston, USA) diluted 1: 1000, anti-LC3B antibody (Abclonal, Wuhan, China) diluted 1: 2000, anti-GAPDH antibody (abcam, Cambridge, MA, USA) diluted 1: 10,000, anti-NLRP3 antibody (Abclonal, Wuhan, China) diluted 1: 2000, anti-IL-18 antibody (Abclonal, Wuhan, China) diluted 1: 2000, and anti-Caspase-1 antibody (Proteintech Group, Inc, Wuhan, China) diluted 1: 2000 for at least 16 h