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. Author manuscript; available in PMC: 2023 Sep 1.
Published in final edited form as: Nat Struct Mol Biol. 2023 Jan 9;30(3):370–382. doi: 10.1038/s41594-022-00887-4

Figure 7. KAKA virions produced in IP6-low cells have fewer and less-stable capsids.

Figure 7

(A) Cryo-ET comparison of KAKA and WT virus produced in IPMK KO + FM1 cells. Tilt-series were collected and reconstructions performed to assess capsid morphology. A total of 48 WT and 24 KAKA particles were analysed. Virions were classified into the indicated categories: Immature (pink), Mature Conical (dark blue), Mature Tubular (light blue), Mature Irregular (green). Slices through representative tomograms of the virions are shown together with quantification. Scale bars, 100 nm. (B) TIRF microscopy on WT virions produced in 293T cells and KAKA mutants produced in IPMK KO + FM1 cells. Virions were adhered to Ibidi slides and treated with SLO in the presence or absence of IP6. Samples were fixed, permeabilised and labelled with VSV-G and p24 antibody. Virions from three independent images for each condition were analysed for mean fluorescence object intensity above threshold (see Supplementary Figure 8). A one-way ANOVA was used for statistical analysis and significant differences indicated [P = 0.01 (**), < 0.0001 (****)]. Right panel shows representative images of virions used in the analysis. Scale = 20μm.