Indoxyl Sulfate Induces Renal Fibroblast Activation through a Targetable Heat Shock Protein 90-Dependent Pathway
Effects of IS on the NRK-49F rat kidney fibroblast cell line. (a) Collagen I mRNA and protein in IS-treated and untreated NRK-49F cells. Data are expressed as fold change versus untreated cells ( and , respectively). Photos are representative of collagen I expression and pattern evaluated by immunocytochemistry (magnification ×400). (b) α-SMA protein expression in IS-treated and untreated NRK-49F cells. Data, obtained by western blot analysis, are expressed as fold change versus untreated cells (). Pictures are representative of α-SMA expression evaluated by western blot and immunocytochemistry. (c) TGF-β and (d) MCP1 mRNA and protein levels in IS-treated or untreated NRK-49F cells. mRNA expression is evaluated by real-time PCR, normalized for GAPDH mRNA. Western blot analysis is normalized for β-actin; values are expressed as fold versus untreated cells (). (e) Flow cytometry analysis of the proliferation index, after 48 hours of exposure, and ROS production, after 60 minutes of exposure, as detected by the probes CFSE and CellRox, respectively, in NRK-49F cells treated or not with 20 μM IS. UC: untreated cells; IS: indoxyl sulfate-treated cells.
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