Figure 6

Labeling of Chlamydia-infected cells with 1-NBD-GPC. HeLa cells were grown in T-75 flasks and labeled with 5 µM 1-NBD-GPC (green) for 2 hours at 37 °C. The cells were washed and incubated in fresh medium for 2 hours before infection with C. trachomatis strain D. Live imaging was performed after 36 hours using a Keyence microscope equipped with a 20x objective (set a; bright-field and merged images with NBD fluorescence are shown). Note the presence of labeled and unlabeled cells. Cropped images (bright-field, NBD fluorescence, and merged images) showing 2 infected cells with labeled inclusions and one cell with an unlabeled inclusion are shown on the right (set b). Lipids were extracted from the treated cells and analyzed in the experiments presented in Fig. 7. Labeling experiments were performed twice and shown images were taken during the second experiment.