Skip to main content
. Author manuscript; available in PMC: 2018 Jan 1.
Published in final edited form as: J Hepatol. 2016 Sep 4;66(1):55–66. doi: 10.1016/j.jhep.2016.08.018

Figure 7. ‘Old’ NS5A species are more hyperphosphorylated and are associated with a higher ratio of positive:negative strand genomes.

Figure 7

(A). Schematic of experimental design to isolate ‘new’ and ‘old’ NS5A. Subgenomic replicon cells expressing NS5A-SNAP were either untreated or treated with 5μM SB 220025 and Sblock or STMR was applied at the indicated time points before detergent lysis and immunoprecipitation of TMR-labeled NS5A with anti-TMR antibody.

(B). Cell lysates (left panel) and immunoprecipitated TMR-NS5A (right panel) obtained as described above were subjected to immunoblotting for NS5A and β-actin.

(C). Quantitation of hyperphosphorylated ‘new’ versus ‘old’ NS5A, presented as a percentage of total NS5A; values are means±SEM of 5 independent experiments. ** p<0.001.

(D). Positive and negative strand RNA associated with ‘new’ and ‘old’ TMR-labeled NS5A was quantitated by strand-specific quantitative RT-PCR. Values shown are ratios of positive:negative strand RNA and are means±SEM of 4 independent experiments. * p<0.01.