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. 2016 May 16;113(22):E3101–E3110. doi: 10.1073/pnas.1520255113

Fig. 2.

Fig. 2.

S. aureus rsp mutants are less hemolytic and show altered kinetics of cytotoxicity. (A) Hemolysis by S. aureus is drastically reduced in an rsp mutant but is readily restored to wild-type (WT) levels by expressing rsp in trans (Comp) in both S. aureus backgrounds, 6850 and USA300. Statistical analysis was performed by one-way ANOVA and Tukey’s post hoc analysis. ***P < 0.001. (B) Host cell cytotoxicity assayed at 4 h after infection is significantly reduced in rsp mutants compared with wild-type (WT) and complemented mutants (Comp) in infected HeLa epithelial cells for both S. aureus strains, 6850 and USA300. ni, uninfected control. Statistical significance was determined by one-way ANOVA. *P < 0.05; ***P < 0.001. (C) Mutation within S. aureus rsp delays pathogen-induced cytotoxicity. HeLa cells were left uninfected (control) or infected with S. aureus wild-type (USA300 WT), an isogenic rsp mutant, and a complemented mutant (USA300 Comp) along with mutants within the global regulators agrA and saeR (SI Appendix, Table S1). Kinetics of cytotoxicity were monitored over time by propidium iodide staining and flow cytometry, here depicted on the y axis using a log scale. Statistical analysis at each time point was performed by one-way ANOVA and Tukey’s post hoc analysis. *P < 0.05 (rsp mutant compared with wild-type).