Date of download: 6/21/2016 Copyright © 2016 American Medical Association. All rights reserved. From: In Vitro and Ex Vivo Delivery of Short Hairpin RNAs for Control of Hepatitis C Viral Transcript Expression Arch Surg. 2012;147(4): doi: /archsurg Figure 1. Small interfering RNA (siRNA)–mediated hepatitis C virus (HCV) transcript knockdown in vitro. Clone B cells were transfected with lacZ (control), 5′ untranslated region (272), and NS4B (NS4) siRNAs. RNA was harvested in 3 days. Quantitative polymerase chain reaction for the HCV replicon transcript was performed, and transcript expression in NS4 (light gray bars) and 272 (dark gray bars) relative to lacZ control–transfected cells (black bars) was calculated. Columns represent the mean relative expression from a minimum of 3 independent experiments per time point. Error bars indicate SEM. Figure Legend:

Date of download: 6/21/2016 Copyright © 2016 American Medical Association. All rights reserved. From: In Vitro and Ex Vivo Delivery of Short Hairpin RNAs for Control of Hepatitis C Viral Transcript Expression Arch Surg. 2012;147(4): doi: /archsurg Figure 2. Hepatitis B virus (HBV) and NS4B (NS4) short hairpin RNA (shRNA) expression in vitro. HEK293 cells were transfected with either pAAV-eGFP-HBV or pAAV-eGFP-NS4 plasmids as indicated, and RNA was harvested in 3 days. MicroRNA (miRNA) Northern blotting was performed and the membrane was probed with HBV-specific (A), NS4-specific (B), and miR16-specific (C) radiolabeled probes. Figure Legend:

Date of download: 6/21/2016 Copyright © 2016 American Medical Association. All rights reserved. From: In Vitro and Ex Vivo Delivery of Short Hairpin RNAs for Control of Hepatitis C Viral Transcript Expression Arch Surg. 2012;147(4): doi: /archsurg Figure 3. Short hairpin RNA (shRNA)–mediated hepatitis C virus (HCV) transcript knockdown in vitro. Clone B cells were transfected with either pAAV-eGFP-HBV or pAAV-eGFP-NS4 plasmids as indicated, and RNA was harvested in 3 days. Bright field (A), fluorescence (B), and merged (C) microscopic images are shown from a representative field 3 days after transfection to illustrate typical transfection efficiency. D, Northern blotting of total RNA was performed and the membrane was examined with an NS5B- specific probe for detection of the clone B replicon transcript. 18S RNA in the ethidium bromide stained gel is shown for loading control. E, The NS5B probe detects a transcript specific to clone B cells that is not expressed in the Huh-7 parental cell line. GAPDH indicates glyceraldehyde-3-phosphate dehydrogenase. F, Quantitative polymerase chain reaction was performed after reverse transcription of total RNA. The graph shows HCV transcript expression in pAAV-eGFP-NS4–transfected cells (gray bar) relative to pAAV-eGFP-HBV–transfected cells (black bar) at 3 days and represents the mean expression of 3 independent experiments. Error bars indicate SEM. Figure Legend: