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To observe the distribution of GSDME in cytoplasm, cells were treated as indicated and washed with PBS/T (PBS containing Tween-20, 0.05% v/v), then fixed with 4% paraformaldehyde for 20 min before permeabilized with 0.2% Triton X-100 in 3% BSA for 15 min

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Renal function categories were based on the eGFR as per Chronic Kidney Disease Epidemiology Collaboration

This means staying energized during daylight hours and naturally winding down at night

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