Table 1 Experimental kinetic parameters of the QS response.

3O-C12-HSL concentration	Full data aggregation			Side channel level data aggregation
	10 nM	1 µM		10 nM	1 µM		p value
	10 nM	1st pulse	2nd pulse	10 nM	1st pulse	2nd pulse	Sign. conc	Pulse
Maximal intensity (a.u.)	18.9 ± 12.2	52.3 ± 22.2	55.9 ± 24.0	20.6 ± 10.8	55.7 ± 20.0	62.5 ± 22.2	< 0.001	< 0.001
Fluorescence buildup lag (h)	0.3	0.3	1.8	1.6 ± 0.8	0.4 ± 0.2	2.0 ± 1.0	< 0.001	< 0.001
Fluorescence buildup rate (a.u./h)	2.8	8	8.3	5.3 ± 2.6	10.0 ± 5.3	9.2 ± 5.7	< 0.001	0.185
Fluorescence buildup time (h)	4.7	4.8	4.4	3.0 ± 1.3	5.1 ± 3.1	6.1 ± 3.8	< 0.001	0.018
Fluorescence decay lag (h)	1.3	4.8	5.3	0.4 ± 0.6	3.0 ± 2.8	4.6 ± 3.7	< 0.001	< 0.001
Fluorescence decay rate (a.u./h)	1.5	3.7	3.4	3.3 ± 1.1	4.2 ± 3.5	5.3 ± 5.6	0.001	0.032
Fluorescence decay time (h)	7.7	9.1	9.3	3.5 ± 2.1	10.3 ± 3.0	8.7 ± 3.3	< 0.001	< 0.001

3O-C12-HSL concentration	Full data aggregation			Replicate-level data aggregation
	10 nM	1 µM		10 nM	1 µM		p value
	10 nM	1st pulse	2nd pulse	10 nM	1st pulse	2nd pulse	Sign. conc	Pulse
Quorum buildup lag (h)	2.6	0.7	1.5	2.5 ± 0.7	0.7 ± 0.2	1.0 ± 0.9	0.040	0.576 (0.510)
Quorum buildup rate (1/h)	0.1	0.5	0.4	0.14 ± 0.08	0.5 ± 0.2	0.6 ± 0.1	0.105	0.822 (0.839)
Quorum buildup time (h)	–	1.5	2.1	–	1.5 ± 0.6	1.3 ± 0.3	0.794	0.614 (0.661)
Quorum decay lag (h)	0.8	5.6	5.6	0.2 ± 0.7	6.5 ± 1.6	7.7 ± 2.6	0.012	0.545 (0.270)
Quorum decay rate (1/h)	0.07	0.1	0.07	0.10 ± 0.09	0.12 ± 0.02	0.07 ± 0.02	0.747	0.034 (0.124)
Quorum decay time (h)	–	6.9	10.3	–	6.1 ± 1.5	8.2 ± 2.6	0.080	0.316 (0.164)
Quorum-on duration (h)	0	13.9	13.7	3.5*	14.5 ± 2.2	16.4 ± 3.5	–	0.463 (0.137)

See the “Methods” section and Supplementary Fig. S5c,d for the definitions of the kinetic parameters. Where applicable, mean values and standard deviations are indicated. For the 10 nM signal case, the population-level quorum-on state was only achieved in one experimental replicate. *denotes a quorum-on duration calculated based on this sole replicate. p-values were calculated for comparisons of the 10 nM and 1 µM (first pulse) experiments, as well as the 1st and 2nd pulse of the 1 µM experiments (see “Methods”). For comparing the effect of different signal concentrations on the replicate level, the unpaired t-test was used. Both unpaired and paired t-tests were used for the replicate-level analysis of the effect of subsequent signal pulses. p-values corresponding to paired t-tests are indicated in parentheses. A linear mixed-effects model was used for the side channel-level analysis. Calculations were made using unpaired data to analyze the effect of signal concentration, and paired data was used to analyze the effect of subsequent signal pulses.

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