Fig 4

High levels of EspN transcription are required for dominance in the ΔespM strain. (A) Schematic of the −10 region from the mycobacterial optimal promoter driving espN transcription. Pink residues are mutations in the −7 and −12 positions. (B) Relative qRT analysis of the espN transcript compared to sigA transcript levels in M. marinum. Outliers were identified using ROUT analysis, Q = 0.5%. Statistical analysis was performed using ordinary one-way ANOVA (P < 0.0001) followed by Dunnett’s multiple comparisons test. Significance is shown relative to the ΔespM/pespN or ΔespMΔespN/pespN strain. ****P < 0.0001. Data include three biological replicates each in technical triplicate. (C) sRBC lysis measuring hemolytic activity of M. marinum. Statistical analysis was performed using ordinary one-way ANOVA followed by Dunnett’s multiple comparisons test relative to the ΔespM or ΔespMΔespN strain. ****P < 0.0001. Data include three biological replicates each in technical triplicate.