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

Figure 1. G3BP1 engages cGAS in a condensed state.

Figure 1

  1. Turbidity and bright‐field microscope images of indicated groups (left) and the quantitative analysis of turbidity of samples were measured by absorbance at 600 nm (right). 10 μM cGAS, 5 μM G3BP1, 5 μM BSA and 100 nM dsDNA (60 bp) were used in these assays. n = 3 biological replicates.
  2. cGAMP production assays. Recombinant cGAS (10 μM) and G3BP1 (10 μM) were incubated with 100 nM dsDNA. The produced cGAMP was quantitatively analyzed by liquid chromatography–mass spectrometry/multiple reaction monitoring (LC‐MS/MRM). ND, not detected. n = 3 technical replicates.
  3. Bright‐field microscope images of cGAS (10 μM) with indicated concentrations of BSA or G3BP1 (left) and a quantitative analysis of the total area of condensates (right). n = 3 biological replicates.
  4. Time‐lapse imaging of liquid droplets fusion after mixing 10 μM cGAS‐mCherry with 1 μM FAM‐labeled dsDNA.
  5. Time‐lapse imaging of liquid droplets fusion after mixing 40 μM cGAS‐mCherry with 10 μM G3BP1‐mEGFP.
  6. FRAP analysis of DNA‐induced droplets of cGAS (1 μM dsDNA, 10 μM cGAS‐mCherry), the yellow dotted circle indicated the region of photobleaching.
  7. FRAP curve of (F). n = 3 biological replicates.
  8. FRAP analysis of cGAS‐G3BP1 droplets. 10 μM cGAS‐mCherry and 5 μM G3BP1 were used. The yellow dotted circle indicated the region of photobleaching.
  9. FRAP curve of (H). n = 3 biological replicates.

Data information: Representative images are shown (A, C–F and H). Error bars, mean with s.d. (A–C, G and I), *P < 0.05, ****P < 0.0001, two‐tailed t‐test. NS, non‐significant; FRAP, fluorescence recovery after photobleaching. Scale bars, 10 μm (A and C), 5 μm (D), 3 μm (E), 2 μm (F and H). See also Fig EV1.