Extended Data Fig. 8: Intracellular growth phenotypes associated with the ∆sidJ mutant expressing SdeA and its mutants.
From: Regulation of phosphoribosyl ubiquitination by a calmodulin-dependent glutamylase
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 on the basis of total bacterial counts at the indicated time points and those of the 2-h time 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 isocitrate dehydrogenase and tubulin as loading controls, respectively (right). In each case, results are from one representative experiment performed in triplicate from three independent experiments; error bars represent s.e.m. (n = 3).
