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. 2020 Sep 17;15(9):e0239153. doi: 10.1371/journal.pone.0239153

Fig 5. Treatment of HCV infected cells with Sofosbuvir significantly reduced HCV induced RTN3L&S and cell released infectious exosomes.

Fig 5

(A) Huh7-JFH1 infected and (B) HCV FL-Replicon cells were treated with Sofosbuvir or not as indicated alongside non-treated Huh7.0 control cells. 10-days post-Sofosbuvir treatment, cell culture supernatant, and cells were harvested. (A&B) Total protein was extracted from cells and analyzed by western blotting probing for HCV NS3 and RTN3L&S with β-actin serving as a control for equal protein loading. (C) Cell released exosomes in cell-cultured supernatants were quantified by NanoSight NS300 and normalized per cell based on total cell counts at the end of treatment. (D) Cell released exosomes from treatment conditioning in (B) were co-cultured with naïve Huh7.0 cells (5000 Exosomes: 1 Huh7.0 cell) over 72h. Total RNA was then extracted from cells and analyzed by RT-qPCR for HCV RNA using 18s as a housekeeping gene. Data for 3 repeat experiments are presented with */#p<0.05 considered significant by the Mann -Whitney U test.