Fig. 5: Mechanistic investigation of MAZ/G4 condensate formation.

a In vitro droplet formation assays of mCherry-MAZ WT and indicated mutants. b Effects of CCND1-G4 motifs on the droplet formation of mCherry-MAZ WT and its mutants. c Detection of punctum formation in HL-7702 and HepG2 cells transfected by expression vectors of MAZ WT and indicated mutants. d Detection of punctum formation in HepG2 and SMMC-7721 cells that were infected by lentivirus carrying sh-Cont or sh-MAZ-3’UTR, and transfected by indicated MAZ WT and mutant constructs. e A schematic model of G4-recruited MAZ to the CCND1 promoter to induce its phase separation. G4-ZF2 interaction promotes MAZ phase separation, while the ZF3-5 are responsible for recruiting additional MAZ proteins and coregulators into the phase-separated condensates. In (a, b), quantification of droplets’ numbers and area are shown at right with P-values indicated on top. Data are presented as mean ± s.e.m. (n = 6 fields each). Scale bar: 20 μm. In (c, d), quantification of punctum numbers is shown at right with P-values indicated on top. Data are presented as mean ± s.e.m. (n = 30 cells/each). In each insert, 5× magnification is used and the scale bar is 2 μm. In (a–d), unpaired two-tailed Student’s t-test was used for statistical analysis. Source data are provided as a Source Data file.