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

Figure 3. Determination of cell viability in 420Bla+ HCV stable cells.

Figure 3

(A and B) Huh7 cells were mock transfected or transfected with 420Bla RNA. Twenty-four hours later, cells were selected with (+) or without (−) 10 μg/ml of blasticidin for 4 days. The Mock TF represents mock-transfected Huh7 cells. The “+/−” indicates the 420Bla-transfected cells selected with blasticidin for 3 days and replenished with fresh medium for 1 day. The cells were analyzed for protein expressions (A) and cell viability (B) as described in “Materials and Methods”. (C and D) Huh7 cells were mock infected or infected with 420Bla virus at an MOI of 0.01 for 24 hr. Three days later, cells were selected with (+) or without (−) blasticidin for 4 days. The “Mock IF” represents Huh7 cells without 420Bla virus infection. The “+/−” indicates the 420Bla-infected cells selected with blasticidin for 3 days and replenished with fresh medium for 24 hr. The cells were then analyzed for protein expressions (C) and cell viability (D). (E and F) Huh7 cells were transfected with an pCMV-FLAG-Bla-expressing plasmid or an empty vector. At day 2 post-transfection, cells were selected with (+) or without blasticidin for 4 days. The Vector represents Huh7 cells transfected with empty vector. The “+/−” indicates the pCMV-FLAG-Bla transfected cells selected with blasticidin for 3 days and replenished with fresh medium for 1 day. The cells were assayed for protein expression (E) and cell viability (F). Data represents mean ± SEM (n = 3).