Alignment of Rab5 sequences across different spices. (a) Alignment of the aa sequences of human Rab5A and Rab5 proteins from Rattus norvegicus, Danio rerio, Caenorhabditis elegans, Drosophila melanogaster, Saccharomyces cerevisiae, Rhizoctonia solani, Dictyostelium discoideum, and Leishmania donovani. Identical and conserved substitution residues among all spices are highlighted in red and blue, respectively. Arrowheads indicate conserved lysine residues. (b) HeLa-FcγRII cells expressing mRFP-Rab5A (WT), (K22R), or (K33R) were infected with the WT L. pneumophila Lp01 strain for 1 h at an MOI of 10. After infection, cells were fixed and stained with anti-Legionella antiserum for detection of extracellular bacteria, and then permeabilized and further stained with DAPI for detection of intracellular bacteria. Bar, 5 µm. Arrows indicate the position of intracellular L. pneumophila. Data are representative of three independent experiments (100 vacuoles were scored in each experiment). Results are shown as means ± SD. Values are determined using one-way ANOVA with Tukey’s multiple comparisons. n.s., not significance.