Fig. 4

TRAP IL-1 decreases HP1y foci number in SW48, while increasing it in SW48 CXR. Sensitive and resistant SW48 cell lines (4 × 10⁴ cells per well) were seeded on 13 mm coverslip. Twenty-four hours later, SW48 cells were treated for 4 days with control medium (10% FBS), IL-1 (α and β, 10 ng/ml each), and CTX (50 μg/ml) alone and in combination with TRAP IL-1 (20 μg/ml) (a), while SW48 CXR cells were treated with control medium (10% FBS + CTX 50 μg/ml), and TRAP IL-1 (20 μg/ml) (b). After fixation, permeabilization, and blocking with 2% BSA, cells were incubated with anti-HP1y antibody. The next day, cells were washed, incubated with secondary antibody FITC mouse, and stained with DAPI (nuclei). Finally, coverslips were mounted on slides and examined under a fluorescence microscope. Number of HP1y foci was determined and analyzed through ImageJ software from three different fields for each sample. Histograms shows the average of HP1y foci for each cell. Statistical analysis was carried out using two-way ANOVA with Tukey’s multiple comparisons test ***p < 0.001, ****p < 0.0001. Scale bar, 20 mm