Fig. 5. Effect of varied metabolite supplementations and initial cell densities on growth and population size in three-member co-cultures.

a, Diagram of three-member co-culture AKH_III via one-way communication. b,c, The growth of co-culture (b) and population percentage (c) of each member of co-culture AKH_III with and without exchanged metabolite (em) supplementation (mg l⁻¹) at 48 h. em_0 means no em supplementation; the supplementation (final concentration, mg l⁻¹) of ade, Lys, His and Trp were 2.5, 5, 10; 12.5, 25, 50; 10, 20, 40 and 10, 20, 40, respectively. N = 3 biologically independent samples and data are presented as mean ± s.d. One-way ANOVA, followed by Bonferroni’s multiple comparisons test with 95% confidence intervals were performed using GraphPad Prism 9.5.0 and P values are noted. d, Diagram of three-member co-culture AKW_VI via two-way communication. e,f, The growth of co-culture (e) and population percentage (f) of each member in co-culture AKW_VI with and without em supplementation at 48 h. N = 3 biologically independent samples and data are presented as mean ± s.d. One-way ANOVA, followed by Bonferroni’s multiple comparisons test with 95% confidence intervals were performed using GraphPad Prism 9.5.0 and P values are noted. g, The growth curves of co-culture AKW_VI (OD) and estimated growths of three members including GOD, BOD and ROD at different initial cell densities of OD₇₀₀ 0.067, 0.078, 0.102 and 0.148 in 72 h. OD indicates the total OD₇₀₀ values of the co-culture. GOD, BOD and ROD represent the estimated OD values for GFP-, BFP- and RFP-tagged populations, respectively. The initial ratio was 1:1:1 for each member in these co-cultures. N = 3 biologically independent samples and data are presented as mean ± s.d.

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