Overnight cultures of wild type and mucoid E. coli MG1655 carrying plasmid pBAD18Cm::PrprA-mCherry (reporter for induction of the Rcs pathway (Konovalova et al. 2018 (link)) were grown in LB media. These cultures were diluted 1/1000 in fresh LB and incubated for 30 min with vigorous shaking (300 RPM) at 37°C. We then added GFP-labelled T7 phage (Vidakovic et al. 2018 (link)) at 0.1 MOI and returned to incubation in a stationary incubator at 37°C. The phage free control and phage infected culture slides were sampled at 15–30 min intervals and observed on an inverted fluorescent microscope (Leica DMi8 with LasX Premium software). Images are an overlay of DIC, TRITC and GFP channels, acquired using a 40x objective (total magnification 400x).
Quantification was performed using Image J. Object thresholds were established using DIC images, and particle analysis was performed on all particles exceeding 0.1 µM2 area. Resulting plots show mean pixel intensity in each fluorescent channel for all identified objects.
Quantification was performed using Image J. Object thresholds were established using DIC images, and particle analysis was performed on all particles exceeding 0.1 µM2 area. Resulting plots show mean pixel intensity in each fluorescent channel for all identified objects.
