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The combination of Mg2+ and deletion of lcrQ can fully rescue the ΔrfaL mutant. Wild-type, ΔrfaL, ΔlcrQ, ΔrfaL ΔyopN, and ΔrfaL ΔlcrQ strains were grown at 37°C in buffered DMEM with or without EGTA, CaCl2, or MgSO4 added to the medium. Following incubation, cultures were centrifuged to separate secreted proteins (S) from cells (P). Proteins in both fractions were trichloroacetic acid precipitated and immunoblotted using antibodies against YscD, YopE, YopD, and RpoA.
