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After treatments, the slides were fixed with 4% paraformaldehyde at 37°C for 30 min, permeabilized with 0.2% Triton X-100 (Beyotime Institute of Biotechnology) at 37°C for 20 min, blocked with 5% bovine serum albumin (BSA, Beijing Solarbio Science & Technology Co., Ltd.) at 37°C for 30 min and incubated with primary antibodies against HMGB1 (1:100) and GPX4 (1:100) at 4°C overnight

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Even though this is a more rudimentary coordination of cells than might have been needed in even the earliest metazoans, this life-cycle is dependent on a large number of advanced signaling mechanisms