Extended Data Fig. 2: Representative micrographs, 2D classes and western blot analysis of HIV-1 particles used in this study.

a, HEK293T cells were transfected with pcHIV carrying the indicated Gag cleavage site mutations. Particles were collected at 44–48 h post transfection by ultracentrifugation. Viral proteins were detected by immunoblotting with the indicated polyclonal antisera. Observed cleavage patterns are representative of at least three independent preparations for each variant. Antibodies were detected using a LI-COR Odyssey Clx infrared scanner, using secondary antibodies and protocols provided by the instrument’s manufacturer. Positions of molecular mass markers in kDa are indicated at the left. For gel source data, see Supplementary Fig. 1. b, For each dataset a representative cryo-EM micrograph (from at least 1000 micrographs of virus) is shown, illustrating the overall viral morphology (scale bars: 100 nm). Representative high resolution 2D classes of the Gag layer are also shown, labelled with the features that could be identified, including MA, CA. and NC layers. Schematic illustrations of the processing state of Gag constructs are given for each dataset.