FIGURE 3.

The repeat 3 of the LDLR promoter is responsive to hnRNP K knockdown. A, schematic representation of the wild type, 3 mutants, and 1 deletion of pLDLR234Luc reporters. To perform siRNA knockdown and dual-luciferase assay on 96-well plates, 0.5 μl/well of 2 μm small RNA were mixed with 1.5 × 10⁴ HepG2 cells/well. The next day, 100 ng/well of pLDLR234Luc reporter plus 5 ng of pRL-TK reporter as transfection control were cotransfected into these cells. Two days after transfection with different reporters, cells were lysed. The normalized luciferase activities of various reporters are presented in B, and in C the relative luciferase activity in scrambled siRNA-transfected cells is expressed as 1. The data shown are representative of three separate transfections with similar results.