Figure 2

T7SS substrates contribute to resistance to linoleic acid toxicity. (A) S. aureus USA300 wild-type (WT) and USA300 esxA (ΔesxA) or esxB (ΔesxB) deletion mutants were grown in TSB or TSB supplemented with 80 µM linoleic acid (LA). (B) AUCs of biological replicates grown in TSB + LA as in (A) were calculated and presented as % relative to the WT. Means ± SEM are shown. n = 4. (C) S. aureus Newman WT and Newman esxA (ΔesxA) or esxB (ΔesxB) deletion mutants were grown in TSB or TSB + 40 µM LA. (D) AUCs of biological replicates grown in TSB + LA as in (C) were calculated and presented as % relative to the WT. Means ± SEM are shown. n = 4. (E) Growth curves as described in (A) were done with RN6390 wild-type (WT) and RN6390 essC (ΔessC) or esxC (ΔesxC) deletion mutants. (F) AUCs of biological replicates grown in TSB + LA as in (E) were calculated and presented as % relative to the WT. Means ± SEM are shown. n = 3. (G) Newman WT with the empty pOS1 plasmid (WT pOS1) and Newman esxA mutant with either pOS1 (ΔesxA pOS1) or pOS1-esxA (ΔesxA pesxA) were grown in TSB or TSB + 40 µM LA. Data shown in (A,C,E,G) are representative of at least three independent experiments. (H) AUCs of biological replicates grown in TSB + LA as in (G) were calculated and presented as % relative to the WT. Means ± SEM are shown. n = 4. In (B,D,F,H) *indicates P < 0.05 using a Kruskal–Wallis test with Dunn's multiple comparisons test.