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. 2021 Jun 18;22(8):e52447. doi: 10.15252/embr.202152447

Figure EV4. Pseudotyping of cGAMP‐VLPs with IAV Haemagglutinin (HA).

Figure EV4

  1. Schematic representation of cGAMP‐HA‐VLP and Empty‐HA‐VLP production. HEK293T cells were transfected with plasmids encoding HIV‐1 Gag‐GFP, VSV‐G envelope and IAV HA. cGAMP‐HA‐VLPs were collected from cells co‐expressing cGAS WT and Empty‐HA‐VLPs from cells co‐expressing catalytically inactive cGAS AA.
  2. IAV HA is present in HA‐VLPs. HEK293 cells were infected with decreasing amounts of cGAMP‐HA‐VLPs and Empty‐HA‐VLPs (1/5 serial dilutions starting at 2 µl of VLP stocks per well). Infection was monitored 24 h later by quantifying GFP+ and HA+ cells by flow cytometry. cGAMP‐VLPs were used for comparison.
  3. cGAMP‐HA‐VLPs induce a similar IFN‐I response in infected cells compared with cGAMP‐VLPs. Supernatants from infected cells shown in (B) were tested for the presence of IFN‐I as shown in Fig 1C.

Data information: Data in (B) and (C) are pooled from three independent HA‐VLP productions tested in infectivity and IFN‐I bioassays; mean and SD are shown.