Figure 5.

FUCA1 expression contribute to Chemotherapeutic-induced cell death. (A-D). TetOn-p53wt Saos2 cells were were transfected with independent siRNA directed against FUCA1 prior to doxycycline treatment (DOX) for 48 hours. (A) P53 and Fuca1 expression were assessed by Western blotting and immunoblot against Hsp90 was used as a loading control. Cell death was analyzed by flow cytometry, measuring the percentage of cells with sub-G1 DNA content (n = 3 independent experiement, one way Anova *p < 0.01,***p < 0.0001) (B), Caspase 3 activation using an anti-active caspase 3 antibody (C), and western blotting assessing PARP cleavage (D). (E-H) U2OS E1a were trasfected with 2 different siRNA directed against FUCA1 prior treatment with 20 µM Cisplatin (Cis) (E-F) or 20 µM Etoposide (Etop) (G-H) for 48 hours. Cells were assessed for cell death by flow cytometry measuring sub-G1 DNA content content (n = 2 independent experiement, 6 replicates one way Anova,***p < 0.0001) (E-G) and expression of Fuca1 by western blotting. Actin was used as a loading control (F-H).