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Replica Designer Handbags (a) Coimmunoprecipitation assays in 293T cells transfected with expression vectors for the HCV core protein and Flag Dgat1 or Flag Dgat2 proteins and treated with DGAT1 inhibitor (20 or DMSO. After immunoprecipitation with agarose conjugated to antibodies specific for Flag, the core protein was detected by western blotting with antibodies to the core protein. (b) Coimmunoprecipitation of the HCV core protein with endogenous DGAT1 in Huh7 hepatoma cells transduced with a core expressing lentiviral vector. (c) Coimmunoprecipitation of the HCV core protein with endogenous DGAT1 in Huh7.5 and Huh7 Lunet cells electroporated with EGFP Jc1 RNA. (d) Indirect immunofluorescence of core and endogenous DGAT1 in Huh7 cells transfected with wild type or mutant (SPMT) core expression vectors (scale bar, 10 (e) Coimmunoprecipitation assays in 293T cells transfected with expression vectors for wild type (WT), mutant (SPMT) or truncated (1 core protein and the Flag Dgat1 plasmid. Arrows mark unprocessed (top) and processed (bottom) core protein. (f) Model of HCV core recruitment to DGAT1 generated lipid droplets. The HCV core protein interacts with DGAT1 at the ER and thereby gains access to DGAT1 generated lipid droplets. This step is crucial to recruit HCV RNA replication complexes to ER membranes in the vicinity of lipid droplets and to initiate assembly of progeny virions. designed, performed and analyzed most of the experiments and wrote the manuscript. C. Jr. provided the DGAT1 inhibitor. C. Harris also performed DGAT activity assays, analyzed data and edited the manuscript. C. performed the experiments in human primary hepatocytes. constructed the DGAT1 mutants and performed some of the immunoprecipitation experiments. Jr Replica Designer Handbags.