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. 2013 Jan 24;8(1):e54466. doi: 10.1371/journal.pone.0054466

Figure 3. Concentration-dependent restoration of EC-reduced HCV protein synthesis and RNA replication by extraneous COX-2 expression.

Figure 3

Ava5 cells were transfected with the indicated amounts of the COX-2 expression plasmid pCMV-COX-2-Myc encoding cox-2 for 6 h, followed by either (A) (+)-EC or (B) (−)-EC treatment at a concentration of 75 µM for 3 days. “Mock” indicated transfection of control vector pcDNA4/myc-His-A in the presence of 0.1% DMSO. (a) Cell lysates were subjected to Western blotting with anti-NS5B, anti-COX-2, and anti-GAPDH antibodies to evaluate protein expression levels. (b) Total RNAs were subjected to qRT-PCR to evaluate HCV RNA levels. Error bars represent the SD from three experiments. (C) Reduction of HCV protein synthesis by COX-2 gene knockdown. Ava5 cells were transfected with either different amounts (0.5–2 µg) of the COX-2 shRNA or 2 µg of LacZ shRNA vectors as a control group. After 3 days of incubation, cell lysates were prepared for Western blotting with anti-COX-2 and anti-NS5B antibodies.