Anti kim1

Embryoid bodies or kidney organoids were fixed with paraformaldehyde (4%) in PBS for 1 h. Immunofluorescence was performed as previously described (Morizane et al., 2015 (link)). The primary antibodies used were anti-GATA4 (Santa Cruz, #SC-1237), anti-FOXF1 (R&D systems, #AF4798), anti-β III tubulin (Millipore, #MAB1637), anti-OAT1, anti-OAT3, anti-OCT2, anti-KIM1, anti-γH2AX (Cell Signaling, #2577), anti-nephrin (Progen Biotechnik, GP-N2), anti-LTL, anti-podocalyxin (R&D systems, #1658), and Phalloidin (Molecular Probes, #R-415). Alexa 488, 555, or 647 dye-labeled (Molecular Probes; Invitrogen) secondary antibodies were used for immunofluorescence. For 3D whole-mount immunohistochemistry, kidney organoids underwent an additional clearing process after immunostaining as previously described (Hiratsuka et al., 2019 (link)). Immunofluorescence images were obtained using C1 confocal (Nikon) or Stellaris 8 (Leica).

Carnosine and STZ were purchased from Sigma Chemical Company (St. Louis, MO, U.S.A.). The antibodies specific for anti-TNF-α and anti-β-actin were purchased from Santa Cruz Biotechnology (CA, U.S.A.). The antibodies Anti-KIM-1, Anti-P-P65, Anti-P65 and anti-cleaved caspase-3 were obtained from Cell Signaling (Danvers, MA, U.S.A.). Anti-GNMT, anti-COL-I, anti-α-SMA, anti-fibronectin and anti-E-cadherin were obtained from Abcam Biotechnology (Abcam, Cambridge, U.K.). Anti-SAM antibody (MA00201-50) was acquired from Acris Antibodies, Inc. (San Diego, CA, U.S.A.). Blood urea nitrogen (BUN), creatinine and Periodic acid–Schiff (PAS) Kits were acquired from Jiancheng Bioengineering Institute (Nanjing, Jiangsu, China). A Microalbumin Assay Kit was obtained from Abcam Biotechnology (Abcam, Cambridge, U.K.). One step TUNEL Apoptosis Assay Kit was acquired from Beyotime Institute (Nanjing, Jiangsu, China).