Fig. 6

Western blot analysis of purified SCV. Purified SCVs recovered from THP-1 macrophage infected with either live (SCV) or heat-killed (killed) GFP-expressing Salmonella were subjected to SDS-PAGE and Western blotting using antibodies against the indicated host proteins. Purified SCVs show the presence of both early (Rab5 and EEA1) and late (LAMP-1 and LAMP-2A) SCV markers but not LIMP-II which is present in heat (killed) group a indicating the purified SCVs are devoid of lysosomal contamination. Loading controls consisted of a constant amount of uninfected host cell lysates used as an internal standard for the quantification. b Quantification of the relative amounts of the indicated host proteins present in SCVs purified from macrophage infected with either live or heat-killed Salmonella. Bacterial effectors SopE and SopB were found exclusively only on purified SCVs (c). Signal intensities were determined relative to a constant amount of non-infected cell lysates. The data shown are the means and standard error of at least two, independent experiments. Statistical significance was determined by a two-way Annova, where p > 0.5 is considered non-significant (ns); **p < 0.01; ***p < 0.001