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. 2020 Sep 18;12(9):1044. doi: 10.3390/v12091044

Figure 6.

Figure 6

Inhibition of HCV replication as mediated by the combination of miRNA-expressing pLL3.7 vectors. (A) 2 × 104 cells/cm2 and 1 × 104 cells/cm2 of FLuc-JFH1/RLuc cells were seeded in 24-well plates, and after 24 h were transduced with different lentiviral vectors encoding for a combination of shRNAs/lhRNAs targeting the sequences described in the Material and Methods for 72 and 144 h p.t., respectively. At the indicated time points p.t., cells were lysed and processed for fluorimetric detection of transduction efficiency (GFP) and luminometric detection of cell number (RLuc) and HCV replication (FLuc). The FLuc/RLuc (blue columns) and GFP/RLuc (red bars) ratios relative to cells transduced with the indicated pLL3.7 shRNA/lhRNA lentiviruses at 72 (B) and 144 (C) h p.t., are expressed as a percentage of the mean values obtained for cells transduced with pLL3.7 encoding for a non-targeting shRNA (Scrambled). Data are the mean + standard error of the mean relative to three independent experiments. Adjusted p values from the Turkey multiple-comparison post-test are reported for the FLuc/RLuc ratios relative to cells transduced with each lentiviral particle at 72 (D) and 144 (E) h p.t. Green: p value < 0.05; pink: 0.05 < p value < 0.5; red: p value > 0.5; black: no p value calculated. Lentiviral vectors were as follows: #1, pLL3.7/U6-shCypA; #2, pLL3.7/U6-shHCV321; #3, pLL3.7/U6-shPI4KIIIα; #4, pLL3.7/U6-shScrambled;#5, pLL3.7/U6-shHCV321-H1-shCyp-7SK-shHCV353;#6, pLL3.7/U6-shPI4KIIIα-7SK-shHCV321-H1-shCypA; #7, pLL3.7/U6-shHCV321-7SK-shPI4KIIIα-H1-shHCV353; #8, pLL3.7/U6-shHCV321-7SK-shPI4KIIIα-H1-shCypA; #9, pLL3.7/U6-lhHCV321-CypA; #10, pLL3.7/U6-lhHCV-PI4KIIIα.