FIG 5.

MOBKL1B depletion reduces CypA-independent HCV replication despite lack of MOBKL1B-NS5A interaction. (A) NS5A bearing the DEYN mutation does not bind to MOBKL1B. One-STrEP-tag (OST)-fused NS5A was purified using Strep-Tactin Sepharose from Jc1 NS5A-OST wild-type- or DEYN mutant-transfected Huh-7.5 cell lysates. Left panels, Western blot signals of MOBKL1B and NS5A in the input lysates using anti-MOBKL1B and anti-NS5A antibodies. Right panels, Western blot signals following affinity purification of NS5A. (B) Depletion of endogenous CypA or MOBKL1B protein in CD81 knockdown Huh-7.5 cells was analyzed by Western blotting; β-actin is included as a control. (C) Viral genome replication following CypA or MOBKL1B knockdown. At each indicated time point, released luciferase was measured to analyze viral RNA replication (mean of n = 3; error bars, SD). RLU, relative light units.