Extended Data Fig. 8. Intracellular growth phenotypes associated the sidJ mutant expressing SdeA and its mutants.

a. Intracellular defects of the L. pneumophila ∆sidJ mutant can be complemented by SidJ expressed from a multicopy plasmid. The indicated strains were used to infect A. castellanii at an MOI of 0.05 and the growth of the bacteria was evaluated at 24 h intervals. Fold growth was calculated based on total bacterial counts at the indicated time points and those of the 2 h point.

b. Overexpression of a SdeA mutant defective in substrate recognition inhibits intracellular growth of the ∆sidJ mutant. Intracellular growth of the indicated L. pneumophila strains in A. castellanii was evaluated as described in a. In each panel, the expression of SidJ, SdeA and its mutants in bacterial cells and their translocation into infected cells was determined by immunoblotting from total bacterial cell lysates and the saponin-soluble fraction of infected cells, with ICDH and tubulin as loading controls, respectively (right panels). In each case, results are from one representative experiment done in triplicate from three independent experiments; error bars represent s.e.m. (n = 3).