Lectin-based Detection of T-antigen

The T-antigen was evaluated in FFPE tissues from pre-malignant and CRC lesions. Briefly, FFPE tissues were prepared as previously described by us [59 (link)]. After deparaffinization, rehydration, and antigen retrieval, they were incubated with FITC-labeled PNA lectin (1:1000; Vector Laboratories, Newark, CA, USA) for 1 h at room temperature (RT). T24 WT cells were used as negative controls, whereas T-antigen-positive T24 GCNT1 KO cells were used as positive controls [60 (link)]. 4′,6′-diamidino-2-phenylindole dihydrochloride (DAPI, 2.3 × 10⁻³ µg/µL; Thermo Scientific, Waltham, MA, USA) was used as nuclear counterstain. All fluorescence images were acquired on a Leica DMI6000 FFW microscope using Las X software (version 3.0) (Leica, Wetzlar, Germany).