Dm4000 microscope

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.

Formalin-fixed and paraffin-embedded lung tissues were cut into sections of 3 um in thickness for immunohistochemistry (IHC). 4% paraformaldehyde-fixed lung tissues were cut into sections of 6 um in thickness for immunofluorescence (IF) staining. IHC and IF were performed as previously (31 (link)). The primary antibody used were SDF4 (1:100 dilution in 3% BSA), ATF6 (1:200 dilution in 3% BSA), GRP78 (1:200 dilution in 3% BSA) and CHOP (1:200 dilution in 3% BSA). Images were acquired using Leica DM4000 microscope and Nikon A1 Ti confocal microscope.