Cellsens entry system

Morphological changes in IPEC-J2 cells after the treatments were observed under an optical microscope (Olympus, Tokyo, Japan) at a magnification of 100x. Images were acquired using the cellSens Entry system (Olympus).

Five-micrometer-thick sections from formalin-fixed, paraffin-embedded mouse tissues were immunostained after antigen retrieval by incubation for 20 min at 100°C in 0.01 M sodium citrate buffer, pH 6.0, essentially as described previously (Szabo et al., 2016 (link)). Briefly, the sections were blocked with 3% bovine serum albumin (Fraction V, MP Biomedicals, Solon, OH, USA) in PBS and incubated overnight at 4°C with primary antibodies (see Table S3 for further information on antibodies). Bound antibodies were visualized using biotin-conjugated anti-rabbit or anti-goat secondary antibodies (BA-1000 and BA-9500, respectively, Vector Laboratories) and a Vectastain ABC Kit (Vector Laboratories), using 3,3′-diaminobenzidine as the substrate (Sigma-Aldrich). All microscopy images were acquired on Olympus BX43 microscope using an Olympus DP74 digital camera with cellSens Entry system (all Olympus).

Five µm thick sections from formalin-fixed, paraffin-embedded mouse tissues were immunostained after antigen retrieval by incubation for 20 min at 100°C in 0.01 M sodium citrate buffer, pH 6.0 essentially as described previously (Szabo et al., 2016 (link)). Briefly, the sections were blocked with 2.5% bovine serum albumin (Fraction V, MP Biomedicals, Solon, OH, USA) in PBS and incubated overnight at 4°C with goat anti-hEPCAM (R&D Systems, Minneapolis, MN, USA), rabbit anti-hTROP2 (ab214488, Abcam, Cambridge, MA, USA), or rabbit anti-hClaudin-7 (Life Technologies, Rockford, IL, USA) primary antibodies (see Table S2 for further information on antibodies). Bound antibodies were visualized using biotin-conjugated anti-rabbit, or anti-goat secondary antibodies (Vector Laboratories, Burlingame, CA, USA) and a Vectastain ABC Kit (Vector Laboratories), using 3,3′-diaminobenzidine as the substrate (Sigma-Aldrich, St. Louis, MO, USA). All microscopic images were acquired on Olympus BX43 microscope using Olympus DP74 digital camera with cellSens Entry system (all Olympus, Melville, NY, USA).