Fig. 6

NOSTRIN attenuates self-renewal ability of colon cancer cells and promotes inhibitory CDK phosphorylations. A Representative photomicrographs of colonospheres, (magnification 100X, scale bar = 100 μm) formed by control and NOSTRIN over-expressing HCT116 cells. B Quantification of number of colonospheres formed by control and NOSTRIN over-expressing HCT 116 cells. C Immunoblotting for cancer stem cell markers CD133, CD44 and EpCAM using proteins from control and NOSTRIN over-expressing HCT 116 cells. Full length blots are shown in Fig. S3. D Quantification of blots in C using using NIH Image J software. GAPDH was used as endogenous control. E BrdU incorporation assay scoring cell proliferation in control and NOSTRIN over-expressing HCT116 cells. F Immuno-precipitation (IP) with anti-CDK1 antibody followed by western blotting using anti-NOSTRIN antibody in control and NOSTRIN over-expressing HCT116 cells. Full length blots are shown in Fig. S3. G Western blot analysis of p-T14-CDK1, p-Y15-CDK1 and CDK1 using control and NOSTRIN over-expressing HCT 116 cells. GAPDH was used as an endogenous control. Full length blots are shown in Fig. S3. H Quantification of the phosphoproteins (T14 & Y15) relative to basal CDK1 levels from blots in G using NIH Image J software. Full length blots are shown in Fig. S3. Error bars represent standard error of mean from three independent biological replicates. ***p < 0.001