Journal: Scientific Reports
Article Title: Spatial mapping of influenza and coronavirus receptors in the respiratory and intestinal tract epithelium of beef cattle using advanced PixF image analysis
doi: 10.1038/s41598-025-28429-0
Figure Lengend Snippet: Coronavirus receptors in the respiratory tract and intestinal tracts of beef cattle. Confocal images showing angiotensin-converting enzyme 2 (ACE2, A ), transmembrane protease serine 2 (TMPRSS2, B ), aminopeptidase N (APN, C ), dipeptidyl peptidase 4 (DPP4, D ), and carcinoembryonic antigen-related cell adhesion molecule 1 (CEACAM1, E ) expression following immunofluorescence staining. Primary antibodies, ACE2 (4 µg/ml, Cat# sc-390851), TMPRSS2 (0.2 µg/ml; Cat# sc-515727), APN (0.16 µg/ml; Cat# sc-166105), CEACAM1 (4 µg/ml; Cat# sc-166453), and DPP4 (1:50 dilution; Cat# BOV2078, Washington State University Monoclonal Antibody Center, Pullman, WA, USA); Secondary antibody, AffiniPure™ Donkey Anti-Mouse IgG (H + L) conjugated Alexa Fluor ® 488 (15 µg/ml; Cat# 715-545-150). Blue represents nuclear staining using DAPI; scale bar = 20 µM. Representative images of coronavirus receptor expression across tissues are shown. The epithelial lining of the trachea ( Ai ), alveoli ( Aiv ), small intestine ( Av ), and large intestine ( Avi ) showed strong ACE2 expression compared to the bronchus ( Aii ) and bronchiole ( Aiii ). The respiratory epithelium showed extensive TMPRSS2, APN, and DPP4 expression with a multifocal expression on the alveoli ( Bi – iv , Ci – iv , Di – iv ). TMPRSS2 ( Bv – vi ) and DPP4 ( Dv – vi ) were detected uniformly on villi and crypt regions in the small and large intestine epithelia, while APN levels were comparatively low in both intestines ( Cv – vi ). With the exception of the bronchus ( Eii ) and bronchiole ( Eiii ), the levels of CEACAM1 were low to no detection on the epithelia of all the tissues assessed ( Ei , Eiv – vi ).
Article Snippet: Deparaffinized and heat-treated antigen retrieval sections were incubated with Animal-Free Blocker R.T.U (Cat# SP-5035, Vector Laboratories) for 30 min at room temperature, followed by incubation with optimized concentrations of primary antibodies: ACE2 (4 μg/ml, Cat# sc-390851, Santa Cruz Biotechnology, Dallas, TX, USA), TMPRSS2 (0.2 μg/ml; Cat# sc-515727, Santa Cruz Biotechnology), APN (0.16 μg/ml; Cat# sc-166105, Santa Cruz Biotechnology), CEACAM1 (4 μg/ml; Cat# sc-166453, Santa Cruz Biotechnology), and DPP4 (1:50 dilution; Cat# BOV2078, Washington State University Monoclonal Antibody Center, Pullman, WA, USA) overnight (16 h) at 4 °C.
Techniques: Expressing, Immunofluorescence, Staining
