Figure 5

HCV modulates SLC3A2/LAT1 complex to promote cell proliferation. (A) (Upper panel) Huh7.5 cells were transfected with negative siRNA, SLC3A2-specific siRNA or cotransfected with SLC3A2 and LAT1 siRNAs. At 48 h after transfection, cells were either left untreated or treated with 5 mM L-leucine. At 4 h after treatment, GDH activity was measured according to the manufacturer’s instructions. (Lower panel) Cell lysates harvested at 48 after siRNA transfection were immunoblotted with the indicated antibodies. (B) (Upper panel) Huh7.5 cells were either mock-infected or infected with Jc1 for 4 h. At the indicated time points, cells were either left untreated or treated with 5 mM L-leucine. At 4 h after treatment, GDH activity was measured according to the manufacturer’s instructions. (Lower panel) Cell lysates harvested at the indicated time points were immunoblotted with the indicated antibodies. (C) Huh7.5 cells were either mock-infected or infected with Jc1. At 6 days postinfection, total cell lysates were immunoblotted with the indicated antibodies. (D) Huh7.5 cells infected with either mock or Jc1 for 6 days were treated with BCH for 2 h. Total cell lysates were immunoblotted with the indicated antibodies. (E) Huh7.5 cells were treated with either control vehicle (NH₄OH) or 200 µM of BCH. At 96 h after treatment, cell viability was determined by WST assay. (F) Huh7.5 cells infected with either mock (left panel) or Jc1 (right panel) for 4 days were treated with either control vehicle (NH₄OH) or BCH. At the indicated time points, cell proliferation was analyzed by WST assay. (G) Huh7.5 cells were either mock infected (left panel) or infected with Jc1 (right panel). At 4 days postinfection, cells were transfected with either negative siRNA or siRNA mix targeting both SLC3A2 and LAT1 (double). At the indicated time points, cell proliferation was assessed by WST assay.