Figure 5.

SC-GFP inhibited both the MPA of EsxA-ST and the bacteria-induced hemolysis of sheep RBCs. (a) The liposomes containing ANTS (fluorophore) and DPX (quencher) were incubated with the indicated proteins at either pH 7 or pH 4. The dequenching of ANTS fluorescence was recorded with times. (b) The relative fluorescence intensity change between 30 s and 210 s was calculated for each group. (c) Mm(WT), Mm(EsxA-ST) and Mm(ΔEsxA:B) were incubated with SC-GFP at RT, and then same bacterial amount of the strains with or without SC-GFP treatment were incubated with sheep RBCs to test hemolysis. The sheep RBCs incubated with PBS and Triton X-100 solution were used as the blank and positive control to calculate each strain’s hemolysis percentage. The experiments were replicated for three times, the data in curve graph is presented as mean, while the data in column graphs are presented as mean ± SD. For liposome leakage assay, statistical analysis was performed with One-way ANOVA method, followed by Holm-Sidak multiple comparison. For hemolysis assay, statistical analysis within each strain was performed with t-test. Statistical analysis between Mm(EsxA-ST) and Mm(ΔEsxA:B) was performed with One-way ANOVA method, followed by Holm-Sidak multiple comparison. *< 0.05, **P< 0.01, ***P< 0.001