FIGURE 3.

Comparison of release of PSMα and the cytoplasmic proteins FbaA and GAPDH. (A) (Right) PSMα1 expression of HG001, Rom^R and ΔfarE (Rom^RΔfarE) were determined by Real-Time PCR. The strains were aerobically cultured in TSB medium for 16 h. The pellets were used for total RNA extraction. qPCR experiments were carried out to determinate psmα1 and gyrB was used as housekeeping gene. Fold changes were calculated using ΔΔCt method and relativized to the HG001 expression. Error bars indicate standard error. Statistical significances between mutant clones Rom^R, ΔfarE (Rom^R ΔfarE) 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. (Left) Release of PSMα1 into the supernatant of HG001, Rom^R clone and Rom^RΔfarE cultured in TSB for 16 h was determined by HPLC; the relative amount of PSMα was calculated by comparing the peak-area in the samples. All experiments were performed in triplicate. Error bars indicate standard error. Statistical significances between mutant clones Rom^R, ΔfarE (Rom^R ΔfarE) with the wild type HG001 were analyzed by Student t-test: not significant P > 0.05, ^∗P < 0.05, ^∗∗P < 0.01, ^∗∗∗P < 0.001. (B) Comparative release of FbaA (aldolase) and GAPDH (glyceraldehyde-3-phosphate dehydrogenase) over time in HG001, Rom^R clone and Rom^RΔfarE clone. Only TSB was used as control.