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. 2016 Oct 24;291(49):25692–25705. doi: 10.1074/jbc.M116.745471

FIGURE 1.

FIGURE 1.

VHL is a direct downstream target of FOXO3a. A, schematic of VHL promoter constructs. B, promoter reporter activities were measured after transient transfection with a Myc empty vector or Myc-FOXO3a expression vector in HEK293T cells. C, one suboptimal FOXO DBE, TTGTTAC (bold and underlined), was localized to the −567 to −561 region of the VHL promoter. D, compared with the wild-type VHL promoter reporter (−599/+66), the mutant VHL promoter reporter (single G-to-C substitution in the putative FOXO DBE) was not up-regulated by overexpression of FOXO3a and FOXO3a-3A in HEK 293T cells. E, expression of FOXO3a-3A-ER was confirmed by Western blotting assays. F, the mutant VHL promoter reporter (single G-to-C substitution in the putative FOXO DBE) was not up-regulated by FOXO3a activation. HEK 293T cells that constitutively express FOXO3a-A3-ER fusion protein were transfected with the wild-type or mutant VHL promoter reporters in the absence or presence of 4-HT (500 nm).