In vivo–passaged L. monocytogenes, strain EGD-e, serotype 1/2a, and the isogenic deletion mutants Δhly and ΔprfA (49) were cultured in brain heart infusion (BHI) medium until mid-log phase. Heat killing of L. monocytogenes by incubation in phosphate-buffered saline (PBS) for 30 min at 70°C was verified by plating on blood agar plates. B. subtilis subsp. spizizenii were cultured in BHI medium, and E. coli K12 DH5-α were cultured in lysogeny broth (LB) medium.
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