Calnexin 10427 2 ap

The concentration of exosomal proteins was detected using the bicinchoninic acid (BCA) Protein Assay Kit. Based on the quantitative results, the exosome supernatant was denatured in 5 × sodium dodecyl sulfonate (SDS) buffer in proportion and subjected to WB analysis (10% SDS–polyacrylamide gel electrophoresis; 50 µg protein/lane) using rabbit polyclonal antibody CD9 (60232-1-Ig, Proteintech, Rosemont, IL, USA), TSG101 (diluted 1:500; cat. no. abs115706; Absin Bioscience Inc., Shanghai, China), CD63 (sc-5275, Santa Cruz, CA, USA), and calnexin (10427-2-AP, Proteintech, Rosemont, IL, USA).

Formalin-fixed paraffin-embedded sections (2 μm) were used for hematoxylin-eosin (HE), periodic acid schiff (PAS), masson staining, immunohistochemistry (IHC), and immunofluorescence (IF) staining 35 (link), 36 (link). A semiquantitaive histologic scoring was performed to evaluate the degree of glomeruli sclerosis and tubular interstitial damage as before 37 (link). The antibodies used for IHC were MFN2 (9482, CST), DRP1 (ab184247, Abcam), MAPK1 (ab32081, Abcam), and PACS-2 (19508-1-AP, Proteintech). The antibodies used for IF were Calnexin (10427-2-AP, Proteintech), MAPK1 (sc-1647, Santa Cruz Biotechnology), PACS-2 (19508-1-AP, Proteintech), DRP1 (8570, CST), TOM20 (HPA011562, Sigma-Aldrich) and DAPI (D1306, Invitrogen). At least three fields of each section were randomly acquired using a Leica DM4000 microscope and a Nikon A1 confocal microscope and analyzed using ImageJ software.