The plasmid library was used to transfect three different human cell lines: HEK293 (embryonic kidney), HepG2 (liver carcinoma), and HeLa (cervical cancer). Cells were seeded at a density of 0.9 × 105 cells per well into 24- well plates. The cells were transfected 24 h after plating with 500 ng/well of the pIS-0 plasmids. Ten ng of pGL4.74, a Renilla luciferase reporter plasmid, was added to each well as a transfection control. Transfection was performed using 50 μL transfection-mix in Opti-MEM® (Life Technologies, Carlsbad, CA, United States) containing 1.5 μL Lipofectamine® 3000 (Life Technologies, Carlsbad, CA, United States) per the manufacturer’s instructions. Opti-MEM® and culture media were used with no antibiotics.
