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Application should be so light that it just

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Lipopolysaccharide mediates time-dependent macrophage M1/M2 polarization through the Tim-3/Galectin-9 signalling pathway

This procedure involved removing the supernatant and resuspending the mitochondrial pellet in an isolation buffer of 250 mM sucrose, 0.5 mM EDTA, and 10 mM Tris-HCl to achieve a final protein concentration of 20-25 mg/ml [2]