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. 2018 Sep 18;14(9):e1007284. doi: 10.1371/journal.ppat.1007284

Fig 8. Reduced NS3 incorporation into detergent-resistant membranes (DRM) in lipin1-deficient cells.

Fig 8

Huh-7 cells were transduced with lentiviral vectors expressing control or LPIN1-specific shRNAs. HCV polyprotein was expressed in the different cells by vaccinia T7 infection and pTM-NS3/5B plasmid transfection at day 7 post-transduction. Sixteen hours post-transfection, cells were lysed in TNE-0.1% Triton X114 and clear cell lysates were subjected to isopycnic ultracentrifugation in 10–40% sucrose gradients at 120, 000g during 16 hours at 4°C. Gradient fractions were subjected to SDS-PAGE and Western Blot to determine NS3 distribution. (A) Representative Western-Blot of gradient fractions showing the distribution of NS3 in control and lipin1-deficient cells. (B) Quantification of the total and DRM-associated NS3 in the different cell lines, including a control of shControl cells treated with 100nM daclatasvir (shControl+DCTV). Data are expressed as the relative value as compared with the control shRNA. Data are shown as mean and SD of four independent experiments (n = 4). Statistical significance was determined using Student´s t-test (*p<0.05; **p<0.01).