Fig. 2.

Western blot detection of T3SS proteins fused to the Flag epitope in Edwardsiella ictaluri whole-cell lysates. Strains carrying the fusions were cultured in MM19 at pH 7.0, MM19 at pH 5.5, and MMP at pH 5.5. Proteins carrying the Flag epitope were labeled with anti-Flag antibody followed by HRP-conjugated goat anti-mouse antibody to detect EsrC, EseB, and EseG. Detection of EseI required mouse anti-Flag antibody followed by biotinylated goat anti-mouse antibody and HRP-conjugated streptavidin. Loading of equivalent protein amounts across lanes was verified using mouse anti-DnaK antibody, biotinylated goat anti-mouse antibody, and HRP-conjugated streptavidin.