Mouse anti α sma 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).

On day 11 after bleomycin injection, BALF was obtained as previously described [18 (link)] and lungs were harvested for real-time quantitative RT-PCR. On day 21 after bleomycin injection, lungs were harvested for hydroxyproline assay and histological examinations. The degree of pulmonary fibrosis was determined by measuring hydroxyproline content in whole lung tissue as previously described [16 (link)]. After perfusion with PBS, the right lungs were fixed with 4% paraformaldehyde and embedded in paraffin for histological examinations. The sections were stained with hematoxylin and eosin (H&E) or Masson’s trichrome. Furthermore, for detection of α-SMA expression in tissue cells, immunohistochemical staining with mouse anti-α-SMA antibody (1/500 dilution, DAKO, Denmark) was performed using a Histofine Mousestain Kit (Nichirei, Tokyo, Japan). The immunoreaction was visualized by incubation with diaminobenzidine (DAB) chromophoric solution. The sections were then counterstained with Mayer’s hematoxylin.