Volume 146, Issue 3, Pages 812-821.e4 (March 2014) Differential Sensitivity of 5′UTR-NS5A Recombinants of Hepatitis C Virus Genotypes 1−6 to Protease and NS5A Inhibitors  Yi-Ping Li, Santseharay Ramirez, Daryl Humes, Sanne B. Jensen, Judith M. Gottwein, Jens Bukh  Gastroenterology  Volume 146, Issue 3, Pages 812-821.e4 (March 2014) DOI: 10.1053/j.gastro.2013.11.009 Copyright © 2014 AGA Institute Terms and Conditions

Figure 1 Characteristics of HCV genotypes 3a-, 4a-, 5a-, and 6a-specific 5′UTR-NS5A (5-5A) recombinants in transfected Huh.7.5 cells. (A) Schematic diagram of J65′UTR-NS2/JFH17 and HCV 5-5A recombinants. LSG mutations (F1464L/A1672S/D2979G) are indicated. (B) RNA transcripts of 5-5A recombinants with indicated mutations were transfected into Huh7.5 cells, and the percentage of HCV Core and/or NS5A-positive cells was estimated (left y-axis; lines). HCV infectivity titers in culture supernatants at peak infection were determined (mean of triplicate infections ± SEM, right y-axis; bar). J65′UTR-NS2/JFH17 was control. Duplicate transfection experiments performed for these recombinants yielded similar results. See Table 1 for details on transfection and second-passage viruses. Gastroenterology 2014 146, 812-821.e4DOI: (10.1053/j.gastro.2013.11.009) Copyright © 2014 AGA Institute Terms and Conditions

Figure 2 HCV genotype 1−6-specific 5-5A recombinants showed differential sensitivity to protease inhibitors, similar to NS3P recombinants of respective genotypes. Huh7.5 cells in 96-well plates were infected with the 5-5A (A) and NS3P8 (B) viruses, and treated with 7 PIs (Materials and Methods). Experimental control 2a(J6/JFH1)24 was included in each treatment. Control 1a(TN)_FL (full-length)11 was incorporated in (A). Values are means of triplicates in the experiment ± SEM. EC50 for each drug against the different genotype viruses is shown in Table 2. Gastroenterology 2014 146, 812-821.e4DOI: (10.1053/j.gastro.2013.11.009) Copyright © 2014 AGA Institute Terms and Conditions

Figure 3 HCV genotype 1−6-specific 5-5A recombinants showed differential sensitivity to NS5A inhibitor daclatasvir. Cultures infected with 5-5A recombinant viruses were treated with daclatasvir. See legend of Figure 2 and Materials and Methods for details. EC50 is shown in Table 3. Gastroenterology 2014 146, 812-821.e4DOI: (10.1053/j.gastro.2013.11.009) Copyright © 2014 AGA Institute Terms and Conditions