Journal: bioRxiv

Article Title: Transketolase of Staphylococcus aureus is involved in the control of master regulators of stress response during infection

doi: 10.1101/538900

Figure Lengend Snippet: A, B. Intracellular proliferation of USA300 wild type (WT) and Δ tkt mutant (Δ tkt ) strains. Endothelial EA.Hy926 cells expressing mKate2 nuclear restricted red fluorescent protein were infected with GFP expressing WT and Δ tkt strains. One hour after infection, cells were washed several times with gentamicin containing medium to eliminate extracellular bacteria. Gentamicin concentration of 50µg/ml was maintained throughout the experiment. Images were acquired every 30 minutes using Incucyte ® S3 live cell imaging system. A. Representative images (see also video S1 and S2). Red: cell nuclei. Green: GFP expressing bacteria. For each line, a white circle was centred on a unique infected cell. Bar = 100µm. B. Quantitative analysis representing the number of green particles divided by the number of red nuclei during the first day after infection. Blue line: WT strain. Red line: Δ tkt strain. Black line: non infected cells. C. Persistence of intracellular WT and Δ tkt strains. Endothelial EA.Hy926 cells were infected with WT and Δ tkt strains as described above. Gentamicin was maintained throughout the experiment. Survival of S. aureus was assessed by plating and counting CFUs three, seven and ten days after infection. Results are shown as mean ± SD of triplicate measurements.

Article Snippet: Plates were incubated and monitored at 5% CO2 and 37°C for 24 hours in the fully automated microscope Incucyte ® S3 (Essen BioScience).

Techniques: Mutagenesis, Expressing, Infection, Bacteria, Concentration Assay, Live Cell Imaging