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(c) Dose dependent decrease of infectious titers in Huh7


November 30th, 2017

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Replica Bags (a) Infection of shRNA expressing Huh7.5 cells with low concentrations of EGFP Jc1 viral stock. The shRNA numbers correspond to the shRNA targeting sequence in the genes. The available antibodies against DGAT2 do not reliably detect endogenous human DGAT2 in our hands. (c) Dose dependent decrease of infectious titers in Huh7.5 cells transfected with Luc Jc1 or Luc JFH1 viral RNA and incubated with increasing concentrations of the DGAT1 inhibitor or DMSO for 48 h. (d) Dose dependent decrease of infectious virus titers released from freshly isolated primary human hepatocytes infected with HCV Jc1 viral stock and treated with increasing amounts of the DGAT1 inhibitor or DMSO for 3 d. Shown are infectivity titers (in focus forming units (FFU)) expressed as percentage of DMSO control. A single experiment is shown. (e) Real time RT PCR analysis of HCV RNA isolated from cells or from supernatants of Huh7.5 cells electroporated with EGFP Jc1 RNA and treated with DGAT1 inhibitor (20 or DMSO. (f) Western blot analysis of cell extracts described in e lysed at day 4 after transfection. (g) Infection of naive Huh7.5 cells with either intracellular or secreted viral particles isolated from Huh7.5 transfected with Luc Jc1 RNA and treated with the DGAT1 inhibitor (20 or DMSO. LDs, lipid droplets. (i) Western blot analysis of cell extracts or isolated lipid droplet fractions from cells described in h. ADRP, adipose differentiation related protein; CRT, calreticulin; TG, extracted triglycerides analyzed by thin layer chromatography Replica Bags.

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