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. 2006 Jul 26;4:e021. doi: 10.1621/nrs.04021

Figure 2. Fusion of GFP2 or Rluc to ERα and SRC-1 does not alter protein function.

Figure 2

HeLa cells were transfected with 100ng of GFP2-C-ERα (A and B) or GFP2-N-ERα (C and D) expression vector with increasing amounts (100, 200, 400, 600, 800, 1000 ng) of either Rluc-C-SRC-1 (A and C) or Rluc-N-SRC-1 (B and D) expression vector, and 500ng of ERE-luciferase reporter, and the remaining amount of DNA with a vector containing a CMV promoter. The DNA levels were equivalent, with 1600ng of total transfected DNA. To confirm that the fusion proteins had comparable activity to wild type proteins, cells were transfected with reporter gene and 100ng of GFP2-C-ERα or WT-ERα to compare ERα activity (E) or with 100ng of GFP2-C-ERα + 500 ng of Rluc-C-SRC-1 or WT-SRC-1 to compare SRC-1 activity (F). All cells (A-F) were incubated with either vehicle or 10-8M estradiol for 24 hours. The cells were lysed and luciferase expression was measured by a luminometer.