Figure 1

(A) hSef-b suppresses colony formation in TRAMP C2 cells. Cells were stably transfected with 5 µg hSef-b (pCDNA/hSef-b) or an empty vector (pCDNA) along with eGFP construct (0.1 µg) for monitoring transfection efficiency. After one day, transfection efficiencies were microscopically monitored, cells were seeded at different densities and selected with G418 for ~2 weeks. Clones were counted at the end of the selection process. The results are normalized to transfection efficiencies, and are representative of 2 independent experiments. TMTC denotes: too many to count. (B,C) hSef isoforms suppress ERK/MAPK and NF-κB in TRAMP C2 cells. Cells were transfected with Elk-1 or NF-κB- luciferase reporter plasmid along with a control empty vector or with hSef-a or hSef-b expression vector. Cells were treated with FGF2 (2.5 ng/ml) for inducing Elk-1 activation, and with IL-1 or TNF (5 ng/ml) for inducing NF-kB activation. Error bars indicate SEM (N = 2, * p < 0.05). (D,E) hSef-b attenuates IL-1 induced NF-κB (p65) nuclear translocation in TRAMP C2 cells. Control or hSef-b Tet on/TRAMP C2 cells were grown in the absence or presence of dox for 24 hr, then stimulated for 15 minutes with 5 ng/ml IL-1 and immunostained with α-p65 antibody. Representative images (D) were quantified for p65 nuclear localization [E, (N = 2, ** p ≤ 0.004)]. More than 300 cells from each sample were microscopically examined. Bar: 10 µm. (F) hSef-b inhibits cytokine induced NF-κB activation in human cervical carcinoma cells. HeLa cells were transfected with NF-κB luciferase reporter plasmid along with each empty vector (0.5 µg/ml), hSef-a or hSef-b constructs at the indicated concentrations. The assay was performed following stimulation with 5 ng/ml IL-1. FI denotes: Fold Increase.