Effect of Nef on the expression of lipids and the formation of LD.

(A) Formation of LD. RLuc cells cultured on a cover slip were transfected with gfp- or nef.GFP-expressing plasmids, and the transfected cells were stained with ORO (red). (B) Effect of MβCD on Nef-mediated alteration of replicon expression. RLuc cells transfected with nef.GFP-expressing plasmid were treated with the SF-DMEM for 24 hr, followed by a 1 hr treatment with the indicated concentration of MβCD in the SF-DMEM. RLuc activity was then determined in the cells. Means and standard deviation values of triplicate experiments are depicted as bars and lines. (C) Effect of Nef on the promoter activities for the lipid synthesis genes. Each promoter-reporter construct, pGL2B-FAS-1500 (FAS) or LDLRLuc (LDLR), was co-transfected with the indicated amount of gfp- or nef.GFP-expressing plasmids, and at 48 hr post-transfection, a FLuc assay was performed with the cell lysates from the transfected cells using a Firefly Luciferase assay kit.