R9 kim 1 antibody

k-ECM-induced 3D organoids were divided into three groups for nephrotoxicity testing: acetone (1%), cisplatin (0.2 mM) treatment, and control (without chemical reagent treatment) for 3 days, respectively. Increased expression of CYP2E1 and KIM-1 are specific signatures of nephrotoxicity. The cytotoxicity of USC organoids exposed to cisplatin or acetone was assessed using a method described in a previous study,^{3 (link)} using immunofluorescence, as described above. Organoids were exposed to 1% acetone (Fisher, USA) or 200 μmol/L cisplatin for 3 days. Cisplatin was stored as a 200 mM stock solution in dimethyl sulfoxide (DMSO) and diluted to its final concentration in PBS/media. The cisplatin was sterile filtered using a 0.22 mm filter prior to application to the organoids. After 3 days of drug exposure, the organoids were pooled and fixed in 4% PFA. Deparaffinization and H&E staining were performed as described above. Antibodies used: secondary Alexa-488 goat-anti-mouse IgG antibody (1:1000) (Invitrogen, USA), CYP2E1 antibody (1:1000) (Millipore, USA), and R9 KIM-1 antibody (1:500) (Merck & Co, Rahway, NJ, USA). All slides were imaged using an Olympus FV1200 Laser Scanning Confocal Microscope.