Fig. 3: WNT/ß-Catenin signaling pathway is a major target of SARB hybrid treatment.

(A) Representative images of immunofluorescence staining of ß-Catenin in HCT116 cells after treatment for 48 h with SARB 10 and 50 µM. Scale bar—25 µm. The lower panel shows images for the control, SARB 10 µM, and 50 µM group which were digitally enlarged. (B) Representative images of immunofluorescence staining of ß-Catenin in HT29 cells after treatment for 48 h with SARB 10 and 50 µM. Scale bar—25 µm. The lower panel shows images for the control, SARB 10 and 50 µM group which were digitally enlarged. An additional image (derived from another replicate) of Hoechst-stained cells was added to this panel to show the presence of micronuclei when treated with 50 µM of SARB. All images are representative for at least two independent experiments. (C) Western Blot analysis of ß-Catenin in HCT116 and HT29 cells treated with different concentrations of SARB for 48 h. The star (*) indicates that the GAPDH blot is the same as in Fig. 5f. (D) Western Blot analysis of ß-Catenin in SW620 and SW837 cells treated with different concentrations of SARB for 48 h. (E) Western Blot analysis of ß-Catenin in HCT116 and HT29 cells treated with 35 µM of Combi or SARB for 48 h. The star (*) indicates that two other treatments were excluded from this blot. The entire original blot is given in supplementary information. Western Blot images are representative for at least two independent experiments. GAPDH immunoblots served as loading control. (F) HCT116 cells transfected with TOP-FOP-FLASH and ß-galactosidase either treated with SARB for 24 h or left untreated and the data is shown as a percentage relative to DMSO. The reporter assay results represent the average of two independent transfection experiments. RLU relative light units