Goat anti mouse igg igm

Human lung single-cell isolation and flow cytometry analysis were performed as described previously (10 (link), 13 (link)). In brief, human lung tissues were minced and then digested with 2 mg/mL Dispase II, followed by digestion with 10 U/mL elastase and 100 U/mL DNase I. Finally, cells were filtered through a 100 μm cell strainer and lysed with red blood cell lysis to get single-cell suspensions. Antibody staining was similar in human and mouse cells. Flow cytometry was performed with a Fortessa and FACSAria III flow cytometer and analyzed with FlowJo 10.6.1 software (BD Biosciences). Anti–human CD31 (clone WM59, catalog 303118, RRID AB_2247932), CD45 (clone WI30, catalog 304016, RRID AB_314404), and EpCAM (clone 9C4, catalog 324212, RRID AB_756086) were from BioLegend. SLC39A8 (ZIP8) polyclonal antibody (catalog PA5-26368, RRID AB_2543868) and goat anti–mouse IgG/IgM (catalog A-10680, RRID AB_2534062) were from Thermo Fisher Scientific. HTII-280 (52 (link)) was a gift from the laboratory of L. Dobbs (UCSF). Human SIRT1 monoclonal mouse IgG1 (clone 834918, catalog IC7714S) from R&D Systems was used for intracellular staining. Pirfenidone was from MilliporeSigma (catalog P2116), and nintedanib was from Selleck Chemicals (catalog S1010).

Cryosections were used for the staining. AEC2s were stained with HTII-280 (52 (link)) (gift from the L. Dobbs laboratory) and goat anti–mouse IgG/IgM (Thermo Fisher Scientific, catalog A-10680, RRID AB_2534062). ZIP8 was stained with ZIP8 polyclonal antibody (Thermo Fisher Scientific, catalog 20459-1-AP) and Cy3 AffiniPure Donkey Anti-Rabbit IgG (Jackson ImmunoResearch Laboratories Inc., catalog 711-165-152, RRID AB_2307443).