Fig. 4
Evaluation of the HlFer1 promoter truncates using relative luciferase activity in ISE6 cells enriched with different concentrations of ferrous sulfate. The pmirGLO plasmids containing HlFer1 promoter truncates were transfected to ISE6 cells using a 1:1 Lipofectamine 2000 transfection reagent. The cells were incubated for 2 days, replaced with different concentrations of ferrous sulfate (0, 0.1, 1 and 2 mM) enriched media, incubated for another 4 days, and then tested for luciferase activity using a Dual-Glo Luciferase Assay System following the manufacturer’s protocol