Fig 8.

Dominant negative effects of the 4M and K146E apoE mutants. Huh-7.5 cells in 12-well cell culture plates were transfected with increasing amounts (0.11, 0.33, and 1 μg) of pCMV6-XL5 vectors expressing wild-type apoE, the apoE mutant with arginine-to-alanine mutations at residues 145, 146, 147 and 149 (4M), and the apoE mutant with a lysine to glutamic acid mutation at amino acid residue 146 (K146E), respectively. The total amount of DNA was kept constant at 1 μg using vector DNA. DNA was transfected into Huh-7.5 cell using DMRIE-C reagent (Invitrogen) by following the manufacturer's instructions. At 6 h p.t., the medium was replaced with DMEM containing 10% FBS. At 24 h p.t., Huh-7.5 cells were infected with HCV at an MOI of 5. At 24 h p.i., cell lysates and the culture media were collected for detection of apoE by Western blotting (A) and for determination of infectious HCV titers by IHC (B). Mean values and standard deviations of three experiments are shown in panel B.