Fig. 3: Characterization of 2-input multicellular programs.

Workflow characterization of 2-input multicellular programs (a). After the input program design, single-lineage programs are mixed and the multicellular program is implemented. The program characterization is done by fluorescence measurements by flow cytometry and plate reader. The flow-cytometry analysis allows us to observe the percentage of population ON and OFF for one state. Design and characterization of 2-input multicellular program 2MP1 and 2MP2, by flow cytometry (b, c) and plate reader (d, e), respectively. Both multicellular programs were implemented using two different single-lineage strains. The lineage trees for each program and its corresponding genetic DNA device are represented. The inputs are represented by letters, a for aTc inducing Bxb1 Integrase and b for arabinose inducing Tp901-1 integrase. To implement each program, the strains were mixed in similar proportions, grown for 16 h and sequentially induced with each molecule. Each histogram shows the expression of fluorescent reporters at different induction states. A representative example is depicted here. The bar graph corresponds to the mean value of the fluorescence intensity (F.I.) in arbitrary units (a.u) for each fluorescent channel (G (GFP), R (RFP), and B (BFP)) with linear different scales. All experiments were performed in triplicate three times on 3 different days (data distribution in dot plots in Fig. S7c–f). The error bars correspond to the ±standard deviation of the mean of the three different experiments. The dotted line indicates the negative autofluorescence from control strain. Note that GFP226 has a lower fluorescence intensity than sfGFP, as expected.