Figure 5
Spry1 decreases E1A 13s induced SRE transactivation in different cell lines. (A) HeLa cells, (B) C33A cells and (C) NIH-3T3 cells were co-transfected with the SRE-Luc reporter construct (1 μg) and with plasmids (0.5 μg) expressing E1A13S and/or Spry1 as indicated. Empty expression vectors were added to keep the amount of transfected DNA constant. After transfection cells were left serum-deprived for 24 h and then incubated with 10% FCS in DMEM. Serum stimulation was performed for the indicated times (white, 1 h; cross line, 5 h; horizontal line, 7 h; dots, 9 h). The promotor activity of the reporter gene in the presence of empty vectors was set as 1. (D) Expression of Spry1 and E1A13S in the lysates was confirmed in Western blot assays. Results are mean ± S.E.M. between duplicates and representative for 3 individual experiments.