Fig. 2

ALK inhibition by CZB reduces proliferation of CMS1 cell lines. A Cell survival assessment upon CZB addition by Alamar Blue assays. A panel of four CRC cell lines belonging to different CMSs (LoVo-CMS1, LS1034-CMS2, HT29-CMS3, HCT116-CMS4) was tested with increasing doses of CZB. Data are reported as % of cell proliferation in relation to controls ± SD. IC50 values are shown for each cell line. B Colony forming assays of cell lines in (A) treated with increasing concentrations of CZB. Representative images of treated and untreated wells are shown. Cell viability data are reported as % of wells covered area in relation to controls ± SD. Statistics were calculated by one-way ANOVA test. C Spheroid-forming assays of cell lines in (A) treated with CZB 250 nM and 500 nM. Statistic was calculated by one-way ANOVA. Scale bars: LoVo, HT29, HCT116: 250 μm. LS1034: 200 μm. D BrdU incorporation assays of LoVo and SW48 upon CZB treatment. Representative images show BrdU-positive nuclei in red, while total DAPI-stained total nuclei are visible in blue. Quantifications of BrdU-positive nuclei in relation to total nuclei ± SEM is shown. Two tailed unpaired T-test was applied. Scale bar: 25 μm