FIG. 2.

Involvement of CKB in HCV replication. (A and E) Knockdown of endogenous CKB in SGR-N and SGR-JFH1 cells (A) or HCVcc-infected cells (E). Cells were transfected with siRNA against CKB (siCKB-2) or control siRNA (siCont) and were harvested at 72 h posttransfection. Real-time RT-PCR for HCV RNA levels and immunoblotting for CKB and GAPDH were performed. (B) SGR-N cells were transfected with pCAGCKB-C283S or empty vector, and HCV RNA levels and expression of CKB and CKB-C283S were determined 72 h posttransfection. SGR-N and SGR-JFH1 cells (C) or HCVcc-infected cells (F) were treated with Ccr at various concentrations for 72 h, followed by quantification of HCV RNAs and total cellular proteins. ATP levels (D) were determined after transfection with siCKB-2, pCAGCKB-C283S, or treatment with Ccr for 72 h in SGR-N cells. The ATP levels in the cells transfected with negative control siRNA (left), empty vector (middle), and no treatment (right) were set at 100%, respectively. (F) HCVcc-infected cells were treated with Ccr, and the viral core protein levels in cells (left) and supernatants (middle) were determined at 72 h postinfection. Collected culture supernatants were inoculated into naive Huh-7.5.1 cells after the removal of Ccr. After 72 h, the core proteins in cells were determined (right panel). All data are presented as averages and standard deviation values for at least triplicate samples. *, P < 0.05 against control such as transfection with siCont (A and E) or empty vector (B) or nontreatment (C, D, and F).