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Subsequently, samples were washed overnight at room temperature with washing buffer (0.2% Triton X-100 and 0.5% 1-thioglycerol in PBS) and then cleared for 36 h at room temperature in clearing medium (40% N-methylacetamide, 1.455 g/mL Histodenz, 0.1% Triton X-100, and 0.5% 1-thioglycerol in PBS)