Fig. 5
Observation of HlFer1 (F2) promoter activity using Venus in ISE6 cells via fluorescence microscopy (a) and western blotting (b). The pmirGLO-no promoter-Venus, pmirGLO-HlAct pro-Venus, and pmirGLO-HlFer1(F2)-Venus plasmids (1.5 μg) with 1.5 μl Lipofectamine 2000 were transfected into 1 × 106 ISE6 cells in a 24-well plate. Media enriched with 1 mM ferrous sulfate were used 2 days after transfection. Cells were observed under a fluorescence microscope after 4 days of medium enrichment. Western blotting results are representative data of three separate experiments. Scale-bars: 100 μm