Fig. 3: Chemical-inducible cGAS phase separation in tubes.

A Rapamycin-induced heterodimerization of cGAS-FRB and FKBP-proteins. B Binding affinities between AF488-FKBP-proteins and Cy5-cGAS-FRB with or without 200 nM rapamycin. C Binding affinities between AF488-proteins and Cy5-cGAS with or without 200 nM rapamycin. D 200 nM rapamycin induced phase separation of 5 μM AF488-FKBP-proteins, 5 μM Cy5-cGAS-FRB, and 5 μM 99 bp dsDNA. Scale bar: 5 μm. E The cGAMP production of 0, 400, or 800 nM cGAS-FRB mixed with 100 nM dsDNA in the presence of 100 nM viral tegument FKBP-proteins with or without 200 nM rapamycin. F Binding affinities between AF488-ORF9 mutation proteins and 25 bp Cy5-dsDNA. G The cGAMP production of 100 nM cGAS mixed with 100 nM dsDNA in the presence of 0, 400, or 800 nM ORF9 proteins with or without 200 nM rapamycin. Proteins without FKBP tag(left). Proteins with FKBP tag(right). H Binding affinities between G3BP1 and PCBP1 mutation proteins and cGAS. I The cGAMP production of 100 nM cGAS mixed with 100 nM dsDNA in the presence of 0, 400, or 800 nM G3BP1 wildtype and mutation proteins. J The cGAMP production of 100 nM cGAS mixed with 100 nM dsDNA in the presence of 0, 400, or 800 nM PCBP1 wildtype and mutation proteins. K Binding affinities between AF488-FKBP-G3BP1 or AF488-FKBP-PCBP1 and Cy5-cGAS-FRB with or without 200 nM rapamycin. L The cGAMP production of 100 nM cGAS-FRB mixed with 100 nM dsDNA in the presence of 0, 400, or 800 nM FKBP-proteins with or without 200 nM rapamycin. Data are presented as mean ± SEM (N = 3). The time points of (D) were initiated from the inclusion of rapamycin. Statistical significance was assessed by two-tailed unpaired t test in (B, C, E, and K). Statistical differences in (E) were made relative to the controls without the addition of protein. Statistical differences were represented as follows: ns not significant, *P < 0.05, ***P < 0.001.