For immunohistochemistry (IHC), the monoclonal antibodies (mAbs) were anti-CXCL17 (MAB4207, clone 422208; R&D Systems, Minneapolis, MN, USA), anti-CXCL10 (NB600-1426; Novus, Littleton, CO, USA), anti-CXCL9 (MAB392, clone 49106; R&D Systems) and anti-CEA mAb, clone II-7 (Dako, Glostrup, Denmark). Mouse IgG, ready to use (Dako), served as negative control. Anti-mouse Ig ImmPress enhancement reagents kit was purchased from Vector Laboratories (Burlingame, CA, USA). The substrate used was 3,3′-diaminobenzidine (DAB; Vector Laboratories).
For immunofluorescence, the mAbs were FITC-conjugated anti-epithelial cell mAb BerEP4 (F0860, lot 00059670; Dako) and unconjugated anti-CXCL17 mAb. Alexa Fluor 594-conjugated goat anti-mouse IgG (ab150116, Abcam, Cambridge, MA, USA) was used as secondary antibody. Anti-CEA mAb was used as a positive control for indirect staining, and FITC-conjugated mouse IgG2b (X0959; Dako) was used as a negative control.
For immunofluorescence, the mAbs were FITC-conjugated anti-epithelial cell mAb BerEP4 (F0860, lot 00059670; Dako) and unconjugated anti-CXCL17 mAb. Alexa Fluor 594-conjugated goat anti-mouse IgG (ab150116, Abcam, Cambridge, MA, USA) was used as secondary antibody. Anti-CEA mAb was used as a positive control for indirect staining, and FITC-conjugated mouse IgG2b (X0959; Dako) was used as a negative control.