Fig. 4.

Menadione treatment increases HIF1A ARE activity in multiple cell types. (A) Reporter assays in which the region surrounding the HIF1A ARE (HIF1A^WT), or a construct in which the ARE was mutated (HIF1A^Mut), were cloned upstream of a luciferase reporter gene. HepG2 cells were transfected with each plasmid and treated with either vehicle control (EtOH) or menadione for 24 h. Cells were then assessed for luciferase activity, and all values were normalized to a control reporter plasmid driven by the LDHA promoter region. (B) Same as (A) only for MCF7 breast cancer cell. (C) Same as (A) only for MDA-MB-231 breast cancer cells. P-values for menadione versus vehicle for each reporter construct are represented with asterisks (***p ≤ 0.001; **p ≤ 0.01; *p ≤ 0.05, Welch's t-test).