Fig. 5: blsA promoter–driven luminescence under constant darkness conditions.
From: The critical human pathogen Acinetobacter baumannii exhibits light-regulated circadian rhythms
Luminescence reporter activity of A. baumannii V15 pLPV1Z-pblsA::luc clone 1 (A–D) and 17978 pLPV1Z-pblsA::luc clon B (E–H) populations under DD for 7.5 days. The graph in A shows all traces of A. baumannii V15 pLPV1Z-pblsA::luc clone 1, n = 47, while only rhythmic samples are shown in (B), n = 24. Baseline-detrended, normalized luminescence is shown. Average endogenous period (FRP) of rhythmic populations of A. baumannii V15 pblsA::luc clone 1 (23.9 ± 4.5 h, n = 24), and the time window for period calculation is indicated in the top right of panel (B). A representative experiment from two biological replicates is shown. Individual wells are represented in colored lines. C Rayleigh plots showing the phase of the bioluminescent peak under constant darkness DD conditions for the rhythmic population (DD, black dots: 21.9 ± 4.9 h, n = 24; R = 0.1, for the rhythmic population). Lines represent the average peak phase of pLPV1Z-pblsA::luc activity (mean vectors for the circular distributions) of each group. The length of the vector represents the strength of the phase clustering, while the angle of the vector represents the mean phase. Individual data points are plotted outside the circle. The central circle represents the threshold for p = 0.05. (D) Individual luminescence plots for the rhythmic populations of A. baumannii V15 pLPV1Z-pblsA::luc clone 1. The graph in E shows all traces of 17978 pLPV1Z-pblsA::luc clon B, n = 36, while only rhythmic samples are shown in (F), n = 19. Baseline-detrended, normalized luminescence is shown. Average endogenous period (FRP) of rhythmic populations of A. baumannii 17978 pLPV1Z-pblsA::luc clone B (24.67 ± 4 h, n = 19) and the time window for period calculation are indicated in the top right of B. A representative experiment (biological replica 1) from two biological replicates is shown. Individual wells are represented in colored lines. G Rayleigh plots showing the phase of the bioluminescent peak under constant darkness DD conditions for the rhythmic population (DD, black dots: 12.6 ± 5 h, n = 16; R = 0.1, for the rhythmic population). Lines represent the average peak phase of pLPV1Z-pblsA::luc activity (mean vectors for the circular distributions) of each group. The length of the vector represents the strength of the phase clustering, while the angle of the vector represents the mean phase. Individual data points are plotted outside the circle. The central circle represents the threshold for p = 0.05. (H) Individual luminescence plots for the rhythmic populations of A. baumannii 17978 pLPV1Z-pblsA::luc clon B. The parameters were determined using BioDare2. Bacteria were grown at a constant temperature of 23 °C. n = total number of samples; 1 sample = 1 well of a 96-well plate. n rhythmic = number of samples exhibiting circadian rhythmicity under free-running conditions (see Methods for definition).
