Immunofluorescent Labeling of β-Catenin

Immunofluorescent labeling was performed on cell-bearing coverslips using the method described previously [12 (link)]. The CAL-62 cells on cell-bearing coverslips were treated with 100 μM ART or 0.2% DMSO for 48 h. The working concentration of rabbit anti-β-catenin used for immunofluorescence staining was 1:500. Briefly, the cell-bearing coverslips were washed in phosphate-buffered saline (PBS, pH 7.4), incubated in 3% H₂O₂ for 10–15 min, and further incubated with anti-β-catenin (1:500; Proteintech) overnight in a humid chamber at 4 °C. Lastly, the coverslips were co-incubated with FITC-conjugated goat anti-rabbit IgG (1:100; Proteintech) at 37 °C for 1 h in the dark, sealed with anti-fluorescence quenching sealing liquid (including DAPI; Beyotime Biotech), and imaged using a fluorescence microscope (Ni-U, Nikon, Tokyo, Japan).