FIGURE 4.

LDLR transactivation by hnRNP K is independent of intracellular cholesterol levels. A, 0.5 μl/well of 2 μm small RNA was mixed with 1.5 × 10⁴ HepG2 cells/well on 96-well cell culture plates. The next day, 100 ng/well of pLDLR234Luc reporter plus 5 ng of pRL-TK reporter as transfection control were transfected into these cells. Two days after reporters' transfection, culture medium was changed to MEM containing 10% LPDS or MEM containing 10% LPDS plus cholesterol (10 μg/ml cholesterol + 1 μg/ml 25-hydroxycholesterol), and cells were incubated for 24 h prior to cell lysis for performing luciferase activity assays. B, dual-luciferase analysis was performed as described in Fig. 4A using pLDLR1563Luc. Each value represents the mean ± S.D. of 6 wells per condition. ***, p < 0.001. The data shown are representative of three separate transfections with similar results.