Figure 4.

Heat shock protein 90 (Hsp90)–survivin-2B_75-93 precursor peptide complex localized to static early endosomes within human monocyte-derived dendritic cells (Mo-DCs). (a) Human Mo-DCs were incubated at 37°C with Alexa 594-labeled Hsp90–survivin-2B_75-93 peptide complex for 1 h and then washed and fixed. Organelles were stained with an anti-EEA1 mAb for early endosomes, anti-Rab5 polyclonal antibody for early endosomes, and anti-LAMP-1 polyclonal antibody for late endosomes/lysosomes followed by Alexa 488-conjugated goat anti-rabbit IgG or anti-mouse IgG and were visualized with confocal laser microscopy. Arrowheads indicate colocalization of the internalized Hsp90–survivin-2B_75-93 peptide complex and each organelle. (b) To quantify the percentage of colocalization, a single z-plane of one cell was evaluated. For each protein and organelle combination, a total of 150 cells (50 cells from three independent experiments) were analyzed. Data are shown as means + SEM of three independent experiments. *P < 0.01.