Fig. 4

QW24 decreases the self-renewal of colorectal CICs and migration of colorectal cancer cells. a-c, HCT116 cells were pretreated with indicated concentrations of QW24 for 4 h. The pretreated cells were cultured in unattached 96-well plates (1000 cells/well) to form the sphere as mentioned in the Methods, and the spheres images were taken after 1 week (a). The sphere number (b) and diameter (c) were measured. Scale bars, 100 μm. Data are presented as mean ± s.d. (n ≥ 8); *, P < 0.05; ***, P < 0.001. d, QW24 pretreated HCT116 cells were seeded in unattached 96-well plates (1000 cells/well) and cultured for 1 week. The percentage of wells with spheres was determined and computed with limiting dilution analysis (LDA) to determine the sphere-initiating cell frequency. Frequency and probability estimates were computed using the ELDA software. n.s., Not statistically significant; ***, P < 0.001. e, HCT116 cells were treated with the indicated concentrations of PTC-209 or QW24 for 12 h, and cells were trypsinized, washed with PBS and stained with CD44-PE and CD133-APC antibodies. The stained cells were analyzed using flow cytometry. f, HCT116 cells were treated with 2 μM QW24 and the cells were imaged at the indicated time. Scale bars, 50 μm. g, HCT116, HT29 and CT26 cells were resuspended with 0, 1, 2, 4 μM QW24 in serum-free medium and seeded in transwell chambers for 12 h as descripted in Methods and the migrated cells were imaged and counted. Scale bars, 100 μm. Data are presented as mean ± s.d. (n = 3); **, P < 0.01; ***, P < 0.001