Skip to main content
. 2021 Nov 15;23(1):e53166. doi: 10.15252/embr.202153166

Figure 5. Full‐length G3BP1 binds to cGAS and promotes LLPS of cGAS.

Figure 5

  • A
    Schematic drawing of domain architecture and truncations of human G3BP1.
  • B
    Coomassie blue staining of purified cGAS, full length and truncated G3BP1 recombinant proteins.
  • C
    cGAS‐G3BP1 interaction analysis. Full‐length and truncated G3BP1 was pulled‐down using anti‐Flag M2 beads. The cGAS‐G3BP1 interaction was further analyzed with immunoblotting (IB). 2 μg recombinant full‐length cGAS and 2 μg each of truncated G3BP1 were used in the assay.
  • D, E
    Time‐lapse imaging of DNA‐induced LLPS of cGAS, in the presence of full length or truncated G3BP1 (2 μM) (D). Partition coefficient of D was calculated as the total fluorescence intensity of droplets / bulk fluorescence intensity of background (E). n = 3 biological replicates.10 μM cGAS and 100 nM dsDNA were used in the assay.
  • F
    cGAMP production assays. Recombinant cGAS (10 μM) and full length or truncated G3BP1 (10 μM each) were incubated, in the presence or absence of dsDNA (200 nM). The produced cGAMP was quantitatively analyzed by LC‐MS/MRM. n = 3 technical replicates.

Data information: Representative images are shown (D). Error bars, mean with s.d. (E and F). Scale bars, 10 μm (D). See also Fig EV5.

Source data are available online for this figure.