Fig. 4: Effects of turning a module ON and OFF on the activity of the other module.
From: A synthetic cell with integrated DNA self-replication and lipid biosynthesis
a Schematic of co-active DNArep and PLsyn with an emphasis on metabolic and activity crosstalk effects. b Confocal microscopy images of liposome samples show that different substrate additions trigger a specific module activity. ON and OFF labeling indicates presence (ON) or absence (OFF) of substrates/cofactors for either DNArep or PLsyn. Liposome membrane dye (Cy5) is colored in white, LactC2-mCherry in magenta, and dsGreen in green. Scale bar is 5 µm. c Phenotype scatter plots from SMELDit image analysis (LactC2-mCherry vs. dsGreen) on all biological repeats (n = 3) show only one active module if substrates are omitted for the other one (ON or OFF state). Classified liposome subpopulations are labeled as ROI 1-4 and gated in different colors as in Fig. 3b. Appended percentages are calculated from the pooled dataset including all biological repeats. Scatter plots from the individual repeats can be found in Supplementary Fig. 12. d Phenotype heatmap with gated percentage values for ROIs 1, 2, and 4 calculated across all replicates with active and/or inactive modules indicates minor crosstalk between the activity of the DNArep and PLsyn modules. Percentages for individual repeats can be found in Supplementary Fig. 12. e dsGreen and LactC2-mCherry intensity profiles across gated ROIs are similar under both single or joint-module activity. Each dot represents a SMELDit identified liposome. The number of liposomes is 10,851 (ROI 1, full reaction), 7934 (ROI 1, DNArep), 2964 (ROI 2, full reaction), 1686 (ROI 4, PLsyn –dNTPs), 2606 (ROI 4, PLsyn), 3336 (ROI 4, full reaction). Vertical lines indicate the mean value of each data cluster. Source data are available for this figure in the Source Data file.
