Rabbit anti type 1 collagen antibody

Maraviroc was obtained from Sigma-Aldrich (St. Louis, MO, USA) (for in vitro experiments) or GlaxoSmithKline (Brentford, UK) (for in vivo experiments). Mouse CCL3 was obtained from Peprotech (Rocky Hill, NJ, USA). The following antibodies were used as the primary antibodies for immunohistochemical analyses: goat anti-CCR5 antibody (Santa Cruz Biotechnology, Dallas, TX, USA), rat anti-Ly6G antibody (BD Biosciences, San Jose, CA, USA), rat anti-F4/80 antibody (Serotec, Kidlington, UK), mouse anti-α-SMA antibody (Dako, Glostrup, Denmark), rabbit anti-type I collagen antibody, rabbit anti-CD31 antibody and rabbit anti-EGF antibody (Abcam, Cambridge, UK). The following rat anti-mouse antibodies were used as the primary antibodies for the flow cytometric analysis: anti-CD11b antibody (BD Biosciences), anti-CD25 antibody (BioLegend, San Diego, CA, USA), anti-CD45 antibody (eBioscience, San Diego, CA, USA), anti-F4/80 antibody (eBioscience), anti-Foxp3 antibody (eBioscience), anti-Ly6G antibody (Gr-1) (Tonbo Biosciences, San Diego, CA, USA), anti-MIP-1α antibody (R&D Systems, Minneapolis, MN, USA), anti-CCR5 antibody (BioLegend), and isotype-matched control IgGs for individual rat antibodies (BD Biosciences).

Routine morphological analysis was performed using lung tissue sections stained with hematoxylin and eosin (H&E) and the Masson-Trichrome (MT) method. Immunohistochemical (IHC) staining was performed on paraffin-embedded lung tissue sections. Briefly, paraffin sections were deparaffinized. Then, sections were subjected to heat-induced antigen retrieval (HIAR) in a 10 mM citric acid buffer (pH 6). Then, endogenous peroxidase activity was quenched with 3% hydrogen peroxide in PBS for 20 min and subsequently blocked with 10% goat serum for an additional 20 min. Sections were then incubated with either rabbit anti-club cell secretary protein (CCSP) antibody (EMD Millipore, Billerica, MA), rabbit anti-type I collagen antibody (Abcam, Cambridge, MA), or rabbit anti-claudin-10 antibody (Life Technologies, Carlsbad, CA). Next, the sections were incubated for 30 min with goat anti-rabbit IgG, which was conjugated with horseradish peroxidase (HRP) (Histofine Simple Stain Mouse MAX Peroxidase; Nichirei, Tokyo, Japan). Finally, detection of the target protein was performed using 3, 3’-diaminobenzidine (DAB) reagent.