Three-component phase-separating proteins on the phase-separated
lipid membrane. (a) PRM × 4, GRB2-GST, and LAT sequence peptide
were added from solution to interact with ternary mixture GUVs with
phase separation properties (green and gray for separated domains).
LAT sequence peptide (green) was labeled with Atto488 at the C-terminal
and had a polyhistidine tag at N-terminal, so it spontaneously anchored
itself to the membranes by strongly binding to Ni-NTA lipid. (b–g)
Matching typical images of the Atto488 protein channel and the TexasRed-DHPE
lipid channel of GUVs with multiple-phase domains. Scale bars are
5 μm. Lipid compositions were 42% DOPC, 29% DPPC, 14% cholesterol,
5% DOPS, 10% Ni-DGS, 0.1% TR-DHPE.