HCV increases Netrin-1 mRNA and protein.

A. Virions depletion experiment. Huh7.5 cells were infected by HCV over 3 d, and the supernatant (SN) was collected, depleted of HCV particles by ultracentrifugation, and added to naïve Huh7.5 cells. Levels of HCV RNA (left) and Netrin-1 mRNA (right) were quantified by RT-qPCR. Data are represented as mean ± standard deviation (n = 3). B. HCV increases the association of Netrin-1 mRNA with microsomes (endoplasmic reticulum [ER] membrane)-bound polysomes in Huh7.5 cells. Netrin-1, Glucuronidase (GUS), and phosphomannomutase 1 (PMM1) mRNA were monitored for their partitioning in the cytosolic (free polysomes) and microsome (ER membrane-bound polysomes) compartments according to the infection status of the cells through sequential extractions followed by RT-qPCR. Statistical significance was determined using the Mann-Whitney test, p < 0.05. C. Netrin-1 protein expression is increased in HCV-positive cells. The level of Netrin-1 protein was detected in microsomes in Huh7.5 cells, naïve and HCV infected cells by immunoblotting using 200 μg of protein, anti-Netrin-1 antibody and recombinant Netrin-1 (rec. Net) as a control. Infection (core signal) and loading control (β-actin signals) are shown. The underlying data for panels in this figure can be found in S1 Data.