Figure 2

Orf1B is required for efficient secretion of EseJ, EseG, EseB, EseC, and EseD. A Autoaggregation of E. piscicida strains at 24 h post-subculture into DMEM. B Secretion profiles of E. piscicida strains. Samples of ECP from similar amounts of E. piscicida strains grown in DMEM were separated using SDS-PAGE gel and stained with Coomassie blue. EseJ, EseC, EseB, and EseD are T3SS proteins, EvpI, EvpP, and EvpC are T6SS proteins. C Orf1B is required for efficient secretion of T3SS translocon proteins and effectors. Total bacterial proteins (TBP; left panel) and extracellular proteins (ECP; right panel) from similar amounts of the WT strain, Δorf1B strain, Δorf1B/orf1B strain and ΔesaN strain were probed with EseB, EseC, EseD, EseG, EseJ, DnaK, and EvpC antibodies. EvpC, a protein secreted by T6SS but not by T3SS, was used as a loading control. The immunoblotting data shown are representative of three independent experiments.