FIGURE 4.

Comparison of release of lipids and total FAs into the supernatant. (A) Release of lipids. The S. aureus clones (Rom^R clone, Rom^RΔfarE, and Rom^RΔfarR) were aerobically cultured in TSB medium for 8 and 16 h. The supernatants were filtrated and 100 μl of each sample were mixed with FM5-95 to a final concentration of 5 μg/ml. Fluorescence was measured with a Tecan microplate reader using excitation at 565 nm and emission at 660 nm. All experiments were performed in triplicate. The relative lipids in the supernatant were relativized to the HG001 value. Error bars indicate standard error. Statistical significances between mutant clones Rom^R, ΔfarE (Rom^R ΔfarE), farR [Rom^R (pCX-farR)] with the wild type HG001 were analyzed by 1-way ANOVA: not significant P > 0.05, ^∗P < 0.05, ^∗∗P < 0.01, ^∗∗∗P < 0.001. (B) Comparison of FAMEs (C15 to C20). The FAs in the supernatants of 16 h cultures of HG001, Rom^R clone, ΔfarE (Rom^RΔfarE), and farR [Rom^R (pCX-farR)] were qualitatively and quantitatively analyzed by GC-MS. The FAs from C15 to C20 were assigned different colors. The total FAs for each clone are shown in the inserted graph. Experiments were performed independently in duplicate. Error bars indicate standard error. Statistical significances between mutant clones Rom^R, ΔfarE (Rom^R ΔfarE), farR [Rom^R (pCX-farR)] with the wild type HG001 were analyzed by 1-way ANOVA: not significant P > 0.05, ^∗P < 0.05, ^∗∗P < 0.01, ^∗∗∗P < 0.001.