Effect FH535-N on inhibition of Wnt/β-catenin pathway.

(A) Effect of FH535-N on TOPFlash activation. Huh7 cells were co-transfected with Top-Flash and phRL-TK plasmid. After 5 h of transfection, cells were treated with vehicle, FH535 or FH535-N at the concentration indicated in presence of 10 mM LiCl. Vehicle in absence of LiCl was used as control for basal levels of Wnt/β-catenin activity. Results are represented as mean ± SD, n = 3. #: p ≤ 0.05, *: p ≤ 0.001. (B-C). Effect of FH535-N on expression of β-catenin targets. Protein expression levels of downstream β-catenin targets from Huh7 cells treated with FH535-N for 36 h were determined by western blot analysis (B, left pannel). Densitometry analysis was performed using ImageJ software (B, right pannel). mRNA expression of downstream β-catenin targets from Huh7 cells treated with FH535-N for 36 h were determined by RT-qPCR (D).