Additionally, no significant changes were observed in the genes and in microcolonies disperses in response to one or more compounds derived from host cells. medium does not influence microcolony dispersal. As a negative control for the cell-conditioned medium treatments in Fig 2A, DMEM was treated with protease inhibitor, proteinase K, trypsin, chymotrypsin, DNase I, RNase A, EDTA, EGTA or 95C heat prior to addition to the FAM20. A black horizontal line in the panel represents the 3 h time point when treated control medium was added to preformed microcolonies. Data represent the mean SD of three independent experiments.(EPS) ppat.1006251.s003.eps (740K) GUID:?056916DD-3633-45E5-A914-AA935F314058 S4 Fig: Lactate separates into fraction 2 by reverse HPLC. HPLC fractionation was performed to separate the D-lactate (50 mM) by an acetonitrile and water gradient. Fractions were collected every three minutes for a total of 12 min. Activity of fractions 1, 2, 3 and 4 was examined in an induction assay by addition to FAM20 microcolonies. D-lactate (50 mM) was used as a lactate control. A black horizontal line in the panel represents the 3 h time point when fractions were added to preformed microcolonies. Data represent the mean SD of one experiment, a single sample with three images were acquired per well.(EPS) ppat.1006251.s004.eps (616K) GUID:?87ABCBB4-B76F-4D86-A227-AAC97AF1E6F2 S5 Fig: Lactate quantification in CM from cells grown in the presence and absence of glucose. Lactate quantification was performed to examine the concentration in CM collected from cells grown in the presence (+) and absence (-) of glucose. Data represent the Triptorelin Acetate mean SD of lactate concentration from Triptorelin Acetate samples collected three times independently in duplicate. *p < 0.05.(EPS) ppat.1006251.s005.eps (295K) GUID:?56FDD9DE-0B94-42E4-A74B-B189CD3099BF S6 Fig: Lactate is not Serpinf2 involved in bacterial lysis. FAM20 were resuspended (107 CFU/ml) in DMEM containing 1% FBS. After 3 h incubation, DMEM or DMEM containing L-lactate at a final concentration of 50 mM was added to the suspensions. The bacteria were further incubated for 5 h. The CFU/ml was measured every hour for total of 8 h. One representative growth curve out of two is shown.(EPS) ppat.1006251.s006.eps (573K) GUID:?9B738D1E-06BB-4466-8911-BDBCC93FC4B2 S7 Fig: Affect of CCCP on ATP concentration. Samples were taken 10, 20 and 40 min after addition of DMEM or CCCP to FAM20 microcolonies and the concentration of ATP was measured. Data represent the mean SD of two independent experiments in duplicates.(EPS) ppat.1006251.s007.eps (403K) GUID:?F1BA4429-5082-49B1-8D93-224875332E85 S1 Movie: Microcolony dispersal can be induced by the addition of Triptorelin Acetate cell-conditioned medium (CM). CM from 100% confluent Triptorelin Acetate FaDu cells were collected and sterile filtered. The bacteria were first allowed to form microcolonies in DMEM for 3 h. CM was then added to the microcolonies at a 1:1 volume ratio, and the timing of dispersal was examined with live-cell time-lapse microscopy. The time lapse begins 10 min after the addition of the CM and lasts for 20 min.(MOV) ppat.1006251.s008.mov (3.8M) GUID:?4DC921A3-3633-4501-A36C-40775607BDEC S2 Movie: The addition of DMEM control medium does not induce the dispersal of microcolonies. Bacteria were allowed to form microcolonies in DMEM for 3 h. Non-conditioned DMEM was added as a negative control at a 1:1 volume ratio, and the timing of dispersal was examined with live-cell time-lapse microscopy. The time lapse begins 10 min after the addition of the DMEM and lasts for 20 min(MOV) ppat.1006251.s009.mov (3.7M) GUID:?2FE30A4E-8F64-4E49-856F-CE047FC8C22F Data Availability StatementAll data is available in the manuscript Triptorelin Acetate and in the accompanied supplementary files. Abstract The development of meningococcal disease, caused by the human pathogen microcolony dispersal. We show that direct contact with host cells is not required for microcolony dispersal, instead accumulation of a host-derived effector molecule induces microcolony dispersal. By using a host-cell free approach, we demonstrated that lactate, secreted from host cells,.