Bs 6285r

Lung tissue sections (3 µm) were cut from paraffin-embedded blocks individually. Tissue sections were treated with target retrieval solution (pH6, S169984, Dako, Victoria, Australia) in a Decloaking chamber at 110 °C for 15 min, followed by 3% hydrogen peroxide in water (v/v) (H1009, Sigma-Aldrich, Melbourne, Australia) for 15 min and with protein block solution (serum-free, X090930, Dako, Mulgrave, Australia) for 30 min at ambient temperature before applying the primary antibodies. The sections then were immunohistochemically stained using primary antibodies: ACE2 (1:800 dilution, ab15348, Abcam, Melbourne, Australia), TMPRSS2 (1:250 dilution, bs-6285R, Bioss antibodies, Redfern, Australia), Furin (1:200 dilution, bs-13228R, Bioss antibodies, Redfern, Australia), α-SMA (1:500, M0851, Dako, Mulgrave, Australia) and TGF-β1 (1:2000, ab215715, Abcam, Melbourne, Australia). The tissue slides were incubated in an IHC humidity chamber for 1 h at ambient temperature, followed by peroxidase-conjugated polymer backbone-carried secondary antibodies and visualized by 3,3′-diaminobenzidine (DAB) staining (EnVision™ Detection Systems, Peroxidase/DAB, Dako, Mulgrave, Australia).