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Fig. 7. RILP recruitment to SCVs is controlled by Rab7. (A) Representative confocal micrographs showing distribution of overexpressed GFP-RILP (green in merged image) in an uninfected cell (upper panel) or one infected with S. typhimurium (WT, red in merged image) for 10 hours (lower panel). Scale bar: 5 µm. Dotted lines indicate cell perimeter. (B) Representative confocal micrographs showing distribution of endogenous RILP (red in merged images) in an untransfected cell (upper panel) and a Rab7-T22N-expressing cell (lower panel), infected with S. typhimurium (green in merged images) for 10 hours. Vacuolar membrane integrity was assessed by labelling LAMP-1 (blue in merged images). Scale bar: 2 µm. (C) LAMP-1 association with bacteria in HeLa cells infected for 8 hours with S. typhimurium (white bars) or the sifA mutant (grey bars) in cells overexpressing GFP-RILP or GFP-RILP together with myc-Rab7-T22N. LAMP-1 association with bacteria was assessed by confocal microscopy. Results shown are the mean ± s.d. of at least three independent experiments in which more than 100 bacteria were analysed for each experiment.

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