Zagreb Institute Of Immunology

tinue into stationary phase, even though LasR-independent behaviors do not JNK-IN-7 commence to seem till the onset of stationary phase, between eight and 24 h. On the other hand, most bacterial cells in nature will not be developing in optimal laboratory-like conditions. Instead, pathogens usually form biofilms throughout infection. At each day time points, the luminescence values of your plate had been read with a BioTek Synergy 2 plate reader. Data had been collected applying BioTek Gen5 application. At the exact same time points, static cultures with isogenic backgrounds, bearing an empty reporter and prepared identically, have been taken, vortexed to disperse cells lasR Cells Overproduce Pyocyanin Strain Alias Relevant genotype or description Supply or reference P. aeruginosa MTC1 MTC63 MTC390 MTC498 MTC500 MTC537 MTC556 MTC625 MTC626 MTC628 MTC637 MTC723 MTC725 MTC733 MTC735 MTC737 MTC745 MTC747 MTC755 MTC757 MTC759 MTC772 MTC774 MTC789 MTC790 MTC794 MTC795 MTC797 MTC838 MTC842 MTC949 0007-2 0022-1 0024-5 0029-1 0053-2 0063-6 CF1 CF2 CF3 CF4 CF5 CF6 PA14 PA14 Dphz PA14 DlasR PA14 DrsaL PA14 DrsaL DlasI PA14 DpqsA PA14 DrhlI DpqsA PA14 DlasR DrhlI PA14 DlasR DrhlR PA14 DlasR DpqsA PA14 DlasR attB::CTX-1-aacC1 PA14 attB::CTX-1-PlasB-lux PA14 attB::CTX-1-PphzA1-lux PA14 attB::CTX-1-PhcnA-lux PA14 attB::CTX-1-PrhlA-lux PA14 attB::CTX-1-PrsaL-lux PA14 DlasR attB::CTX-1-PlasB-lux PA14 DlasR attB::CTX-1-PphzA1-lux PA14 DlasR attB::CTX-1-PhcnA-lux PA14 DlasR attB::CTX-1-PrhlA-lux PA14 DlasR attB::CTX-1-PrsaL-lux PA14 attB::CTX-1-lux PA14 DlasR attB::CTX-1-lux PA14 DlasR DrhlR attB::CTX-1-PlasB-lux PA14 DlasR DrhlR attB::CTX-1-PphzA1-lux PA14 DlasR DrhlR attB::CTX-1-PhcnA-lux PA14 DlasR DrhlR attB::CTX-1-PrhlA-lux PA14 DlasR DrhlR attB::CTX-1-lux PA14 DlasR DambB PA14 DlasR DphoB PA14 DlasR DrhlI DpqsA lasR wild type lasR179C.T lasR wild-type lasR557T.G lasR520G.A lasRD350362 R This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study doi:10.1371/journal.pone.0088743.t001 and serially diluted to obtain colony-forming unit counts. The resulting CFU counts were then utilised to normalize the luminescence values of their respective 11967625 reporter strains. and its absorbance at 520 nm was study inside a BioTek Synergy 2 plate reader. For some experiments, pyocyanin was quantified directly in culture supernatants by reading the absorbance at 691 nm as previously described. Sterile medium was used as a blank. Pyocyanin extraction and quantification Pyocyanin was extracted from the supernatants of liquid cultures by adding an equal volume of chloroform and vigorously vortexing. The lower, pyocyanin-containing organic layer was then taken and vortexed with an equal volume of 0.two M HCl. The pink pyocyanin-containing aqueous layer was taken, Cheating experiments For cheating experiments, PA14 cells or mixtures of PA14 or PA14 phz cells with lasR cells have been grown in liquid M9 medium containing 1% cas