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. 2013 Jan 22;8(1):e54866. doi: 10.1371/journal.pone.0054866

Figure 2. Maintenance of 420Bla+ stable cells at a productively replicating state in early stage after transfection.

Figure 2

(A) Huh7 cells were transfected with JFH1 or 420Bla RNAs and one day after transfection, cells were seeded at the indicated densities and selected with 10 μg/ml of blasticidin for 4 days. The survival clones were visualized by crystal violet staining. The right panel represents the immunostaining of NS5A protein in blasticidin-resistant cells. Green: NS5A; Blue: nuclei stained by DAPI. (B) Huh7 cells were transfected with JFH1 or 420Bla RNAs and one day after transfection, cells were selected with or without blasticidin. Total cellular RNAs isolated at the indicated times were analyzed for the intracellular RNA amount by semi-quantitative RT-PCR. (C) Cell lysates from another set of cells as shown in (B) were harvested at indicated times and analyzed for expressions of each indicated proteins by Western blotting. (D) Culture supernatants collected from (B) were determined for the viral infectivity. Data represents mean ± SEM (n = 3) (D).