Fig. 5: Programmable self-organization through branching devices.
a, Schematic of founder yeast differentiating into two CAM-displaying progeny that drive pattern formation. b, CAM screening and fluorescence images of yeast aggregates formed by Nb3–Ag3, Nb1–Ag1, SpyTag–SpyCatcher and dockerin–cohesin pairs; fluorescent BY4741 lacking CAMs serves as a negative control. Scale bars, 100 μm. c, Aggregate size as a function of Nb3:Ag3 cell ratios (mean ± s.d., n = 10 independent images from 3 independent experiments); box plots show median, interquartile range and full data range. d, Phase-separated morphologies in snowflake yeast; heat maps quantify red–green cell contacts (mean ± s.d., n = 8 biological replicates). e, Snowflake yeast expressing Ag3 (red) and Nb3 (green) forms heterotypic aggregates through Ag3–Nb3 interactions, increasing red–green associations (mean ± s.d., n = 9 biological replicates). f, Tri-lineage assemblies formed when blue progeny (Nb1 + Nb3) adhere to both red (Ag3) and green (Ag1) cells; interaction frequencies are quantified (mean ± s.d., n = 6 biological replicates). NaN, not a number. Scale bars, 100 μm (d–f). g, Schematic of the differentiation of CHO-K1 cells into two progeny types driving pattern formation; TM, PDGFRβ transmembrane domain. h, In situ differentiation and patterning from around 100 founder CHO-K1 cells after doxycycline (Dox) and TMP induction, with mCherry localized to cell–cell interfaces. Scale bars, 50 μm (2 h, 24 h and 48 h); 100 μm (96 h). i, CD19–anti-CD19–synNotch-mediated patterning in CHO-K1 cells. j–l, Self-organization of differentiated progeny via heterophilic EGFP–LaG16 (j), mCherry–LaM4 (k) or homophilic cadherin (Cdh6 or Cdh1) adhesion (l). Scale bars, 100 μm (i–l). Panels h–l show representative images from three independent experiments. Statistical significance was assessed by unpaired two-tailed Student’s t-tests; *P < 0.05.
