Rabbit anti sp c

The mice were euthanised and the lungs harvested and fixed with 4% paraformaldehyde diluted in PBS. Paraffin sections (2 μm) were stained with haematoxylin–eosin (H&E), Masson's trichrome and the following primary antibodies for immunohistochemistry: rabbit anti-E-cadherin, rabbit anti-α SMA, rabbit anti-vimentin, rabbit anti-MMP7 (Cell Signalling Technology, Danvers, MA, USA), rabbit anti-SP-C (Hycult Biotech, Uden, Netherlands), rat anti-Podoplanin (MBL, Aichi, Japan), rabbit anti-collagen I (Novus Biologicals, Littleton, CO, USA), rat anti-F4/80 (Bio-Rad Laboratories, Hercules, CA, USA), rabbit anti-CD3 (Genemed Biotechnologies, San Franciso, CA, USA), rat anti-PTPRC/CD45R (Aviva Systems Biology, San Diego, CA, USA), rabbit anti-arginase I (GeneTex, Irvine, CA, USA), mouse anti-peptidyl-citrulline (F95) and rabbit anti-S100A4 (Millipore, Burlington, MA, USA) and mouse anti-Laminin γ2 N-terminal fragment (γ2pf; Funakoshi, Tokyo, Japan). Histofine Simple Stain Mouse MAX-PO secondary antibodies (Nichirei, Tokyo, Japan) and the Opal 4-colour Fluorescent IHC kit (PerkinElmer, Waltham, MA, USA) were used according to the manufacturer's protocol. All images were captured by fluorescence microscopy (BZ-X710, Keyence) and analysed by hybrid cell count (Keyence, Osaka, Japan) or Fiji.

Western blotting analysis was performed using the ECL system. The following primary antibodies were used: rabbit anti-PDGFRα, rabbit anti-PDGFRβ (Cell Signalling Technology), rabbit anti-Pad4 (MBL), rabbit anti-SP-C (Hycult Biotech), goat anti-SP-D (R&D Systems, Minneapolis, MN, USA), rabbit anti-COL1A1 (Boster Biological Technology, Pleasanton, CA, USA), rabbit anti-SP-A and mouse anti-tubulin-α (Santa Cruz Biotechnology, Dallas, TX, USA) antibodies. Statistical analysis of the expression levels of each protein was performed using Fiji.