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Env adaptations increase RT packaging, accelerate reverse transcription, and reduce G-to-A mutation levels.

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posted on 2018-04-20, 17:36 authored by Terumasa Ikeda, Menelaos Symeonides, John S. Albin, Ming Li, Markus Thali, Reuben S. Harris

(A) Representative immunoblots of the indicated proteins in viral particles and producer cells from one experiment. (B) Relative RT packaging into viral particles produced in SupT11-A3G cells. RT packaging levels were quantified based on band intensity and normalized by each p24 of the virions (mean +/- SD of 3 biologically independent experiments). (C) Relative RT activity in viral particles produced in SupT11-A3G cells. RT activity was measured for each viral lysate normalized to p24 levels (mean +/- SD of 3 biologically independent experiments). (D to G) Kinetics of early RT, late RT, 2-LTR circle, and proviral DNA during infection of CEM-GFP cells using viruses originally produced in SupT11-A3G cells (mean +/- SD of 3 biologically independent experiments). (H) G-to-A mutation loads of high-fidelity, high-temperature pol amplicons from CEM-GFP cells infected with the indicated viruses (mean +/- SD of 3 biologically independent experiments with a minimum of 10 sequences or 5,640 bp per condition). Statistical comparisons for data in panels B-H were done using Student’s t test (p-values above each panel in comparison to Vif-null HIV-1; *: p<0.05, **: p<0.01, ***: p<0.001).

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