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. 2014 Nov;88(21):12276–12295. doi: 10.1128/JVI.00970-14

FIG 6.

FIG 6

HCV replication is restored to near completion by some glycosphingolipids in FAPP2 knockdown cells. (A and B) Dox-induced control and FAPP2 shRNA cells were transfected with 10 μg of Luc-JFH1 replicon RNA as described in Materials and Methods. (A) At 4 h posttransfection, 0, 50, or 100 μM LacCer was added to transfected cells, and HCV replication efficiency was determined at 4 h and 48 h posttransfection. (B) Additionally, cell lysates were subjected to immunoblotting with αFAPP2 (1:1,000) or αGAPDH (1:8000) antibody. (C) Control and FAPP2 shRNA cells were transfected with replicon RNA as described for panel A but treated with 0, 50, or 100 μM GlcCer. HCV replication efficiency was determined as described for panel A. (D and E) Control and FAPP2 shRNA cells were transfected with replicon RNA as described for panel A but treated with various concentrations of Gb3 (D) or cholesterol as a low-density lipoprotein (LDL) (E). HCV replication efficiency was determined as described for panel A. The data are representative of three independent experiments with triplicate samples for panels A, C, D, and E. *, P < 0.05 (statistically significant); **, P < 0.01 (very significant); ***, P < 0.001 (extremely significant).