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. 2012 Mar 20;22(6):1003–1021. doi: 10.1038/cr.2012.44

Figure 2.

Figure 2

HRE on the MIF promoter and phosphorylation of SRC-3 are required for the activation by SRC-3. (A) MCF-7 cells were transfected with MIF promoter-regulated luciferase reporter combined with either control vector or with SRC-3 expression vector. The luciferase activity was measured 48 h after transfection and normalized against total input protein. The normalized activity from vector-transfected sample was set as 1. The values represented fold activation over the control and were expressed as means ± SD from three independent experiments performed in duplicate. (B) Wild-type MIF promoter or promoter with mutation at HRE or CRE was co-transfected with expression vector for wild-type SRC-3 into the MCF-7 cells. The luciferase activity was determined as in A. Shown are means ± SD from three experiments performed in duplicate. (C) MCF-7 cells were transfected with the MIF promoter, together with the wild-type SRC-3 or the indicated SRC-3 mutant. The luciferase activity was determined as described in A. A schematic of the phosphorylation sites on SRC-3 and a representative immunoblot analysis were shown. The values are means ± SD from three experiments performed in duplicate.