Figure 5
Impact of MazEF on the ClpC effect on intracellular survival of S. aureus DSM20231 in keratinocytes and endothelial cells. HaCaT (a) and EA.hy926 (b) cells were infected with the S. aureus strains DSM20231 (WT; black bars), PBM001 (clpC; white bars), DSM20231 mazEF (mazEF; transversal dashed bars), and DSM20231 clpC mazEF (clpC mazEF; dotted bars) at a MoI of 100, respectively, and co-cultured for 90 min. Extracellular and adherent bacteria were subsequently removed by washing and lysostaphin/gentamicin treatment, and the infected cell lines cultured for up to 168 h in cell culture medium supplemented with gentamicin. At the time points indicated, cells were removed, lysed by sonication, and surviving bacteria in lysates determined by plate counting. Data are presented as mean + SD (n = 6–8 biological replicates). *P < 0.05; **P < 0.01 (Kruskal Wallis test followed by Dunn’s post-hoc test between WT and mutants at a given time point). TpL/G, time post lysostaphin/gentamicin treatment.
