Fig. 1
From: Derivation and characterization of an HIV-1 mutant that rescues IP6 binding deficiency
Derivation of an IP6-independent HIV-1 mutant. A Passaging schematic. B Sanger sequencing of HIV-1 gag indicating emergence of a single-site revertant. Input or viral RNA was isolated at indicated timepoint, and the gag region was amplified by RT-PCR. C Single cycle infection assay confirming the identified substitution rescues infectivity of K359A. 293T cells were transfected with proviral clones of WT HIV-1NHG or indicated mutant, and 48 h post transfection, supernatant was titrated on MT4 cells. Statistical analysis: Student’s T test. D, E Spreading replication assays in MT4 cells (D) or CEM cells (E). Cells were infected with HIV-1WT or indicated mutant at an MOI of 0.001. 16 h post infection, inoculum was washed away and supernatants were collected at the indicated timepoints. Reverse transcriptase activity was measured in the supernatant samples using SYBR-PERT