Figure 1

HCV replicon cells treated with NIM811 have enlarged lipid droplets. Wild type sub-genomic 1b (sg-1b) replicon, a sg-1b NIMr replicon, Huh7.5 cells infected with JFH1 HCVcc or Huh7 naïve cells were grown in the presence of NIM811 or DMSO vehicle control. Following compound treatment, cells were fixed, permeabilized and stained for lipid droplets using BODIPY 493/503. A representative image of the lipid droplets in cells treated with 2μM NIM811 (10X EC₅₀) for 24h is shown (A). B) sg-1b, sg-1b NIMr and naïve Huh7 cells were treated with increasing concentrations of NIM811 for 48h. Images of lipid droplets were acquired and quantified by the Cellomics ArrayScan VTI HCS reader. The object area of lipid droplets in each cell type averaged from three experiments is graphed ± standard deviation. C) sg-1b replicon cells were treated with indicated compounds for 48h and the size of the lipid droplets quantified as in B. The object area of the lipid droplets are graphed ± standard deviation. BILN2061 is an NS3 protease inhibitor with a replicon EC₅₀ of 2nM. D) sg-1b replicon cells were grown in the presence of cyclosporin A (CsA, 8μM) or sanglifehrin A (SfA, 10μM) for 24hr. Lipid droplets were fixed and stained as in (A). A representative image is shown, where green represents lipid droplets and blue represents the nuclei (DAPI).