(A) Secretion of SdeA, VipA, RalF, LidA was assayed using adenylate cyclase (CyaA) fusions. DotL overproduction was accomplished by expressing dotL from the CyaA fusion plasmid. CyaA fusions were expressed in a wild-type strain (Lp02), a dotA mutant (Lp03), and an ΔicmS mutant (JV1962). These strains were used to infect U937 cells and calmodulin-induced cAMP production was measured via an ELISA assay. cAMP assays were conducted in triplicate. Error bars represent the standard deviation from the mean.
(B) Westerns blots were performed in order to measure levels of DotL and of the control protein ICDH.