Fig. 3

Initial co-culture composition influences activity of a phenazine sensitive strain within tri-culture system in LB. a Schematic of tri-culture system in LB media, where PYO drives activity in phenazine sensitive strain, Population C. b NEB10β pZE-phzAG (Population A) and NEB10β pZE-phzMS (Population B) were co-cultured in LB media. After 5 h of co-culture, 20 µL of an overnight culture of PH04 pSox-LasI (Population C) grown in LB media were added to the co-cultures. After 1 additional hour of culture, the CM samples were taken. An AI-1 reporter assay was used to measure the extracellular AI-1 levels in the culture.⁵ The controls “phzS” and “LacZα” indicate growth of monocultures NEB10β pZE-phzS or NEB10β pZE-lacZα, respectively, prior to addition of Population C. Error bars indicate s.d. of technical duplicates. c and d NEB10β pZE-phzAG (Population A) and NEB10β pZE-phzMS (Population B) were co-cultured in LB media. After 5 h of co-culture, 20 µL of the co-culture was added to 180 µL of PH04 pCT10-pET-DsRedExpress2 or PH04 pCT10-pET-eGFP (Population C), c and d respectively, which were regrown from overnight culture to ~ 0.2 OD₆₀₀ in LB media. After 4 h of additional culture relative fluorescence was measured. The controls “phzS” and “LacZα” indicate monocultures of NEB10β pZE-phzS or NEB10β pZE-lacZα, respectively, in addition to Population C. Background fluorescence was subtracted out from samples by subtracting LacZα value from each sample. Error bars indicate s.d. of technical triplicates