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Coverslips were mounted with Immu-mount (Epredia Shandon 9990402) and imaged on an inverted epifluorescence microscope (Leica DMi8) with a 20x/NA 0.40 objective (Leica HC PL FLUOTAR L CORR PH1)

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Clinical studies and preclinical models suggest that Tim-3-expressing macrophages, whose renal expression correlates with the severity of diabetic nephropathy, induce podocyte injury through the NF-B/Tumor necrosis factor-alpha (TNF-) pathway, with reduced nephrin expression being inversely associated with macrophage activation (34)