Figure 6.

Effect of Calebin A on multicellular proinflammatory TME-induced activation of cancer stem cells. Serum-starved cultures of HCT116 cells in 3D-alginate cultures alone (basal control = co) or co-cultured with fibroblasts and T-lymphocytes (proinflammatory multicellular TME cultures) or co-cultured with fibroblasts and TNF-β (10 ng/ml) (TNF-β-TME) were either left untreated or treated with various concentrations of Calebin A (CA) (1, 2, and 5 µM), as described in Materials and Methods. Immunoblotting of whole-cell lysates from HCT116 cells was performed for anti-CD133, anti-CD44, and anti-ALDH1 in HCT116 cells. β-Actin served as an internal loading control in all experiments. Densitometric values were compared with the control, and *p < 0.05 and **p < 0.01 were considered statistically significant.