Figure 4

Localization of 7 putative inclusion membrane proteins in C. pneumoniae -infected cells. HeLa cells infected with C. pneumoniae AR39 for 96 hrs were immunostained with mouse anti-GST fusion protein antibodies plus a Cy3-conjugated goat anti-mouse IgG (red) and a rabbit anti-chlamydial organism antibody plus a Cy2-conjugated goat anti-rabbit IgG (green) and Hoechst to visualize DNA (blue). Antibodies against the GST-putative Inc fusion proteins detected signals inside the inclusions, overlapping with the chlamydial organisms. In contrast, antibodies against GST-CPn0186, a control Inc protein, showed peripheral labelling of the inclusion membrane (bottom panels). All antibody labelings were removed by preabsorption of the antibodies with the corresponding GST fusion proteins (panels i to p), but not the unrelated GST-fusion protein control (q to x).