(A) Western blot analysis with anti-lateral flagellin (1:1,000) polyclonal antibodies of cytoplasmic fractions of A. hydrophila AH-3 (wild type), mutant AH-4427 (without polar flagellin), AH-3ΔlafA (without lateral flagellin), and AH-5502 (rpoN) (lanes 1, 2, 3, and 4, respectively) of bacteria grown in liquid (TSB) or viscous medium (TSB plus 18% [wt/vol] Ficoll) at 25°C in TSB. (B) Western blot analysis with anti-polar flagellin (1:1,000) polyclonal antibodies of cytoplasmic fractions of the same strains and grown conditions as those described for panel A. (C) Western blot analysis with anti-lateral flagellin (1:1,000) polyclonal antibodies of whole cells (WC) and supernatant before shearing (S) and after shearing (SS) of A. hydrophila AH-3 grown in liquid medium (TSB), viscous medium (TSB plus 18% [wt/vol] Ficoll), or an agar plate (TSA) (lanes 1, 2, and 3, respectively).