Figure 3
Z-stack confocal images of co-cultures, at 48 h.p.i by BCG, moving from the apical (top left) through to basolateral (bottom right) aspect for each panel. (a) Co-cultures of endothelial BPAECs overlaid with the BATII epithelial cell line stained with the ATII marker SFTPC (green), phalloidin (red) and the endothelial marker CD31 (cyan). Moving left to right, a tight epithelial cell layer was observed (red and green fluorescence; green box), giving way to the underlying endothelial BPAEC (red and cyan fluorescence, blue box). (b) At 48 h.p.i, the epithelial BATII layer displayed regions of decimation, as evidenced by bare patches in the epithelial layer (red box); however phalloidin staining was still present underneath, in co-localisation with CD31 (blue box), suggesting that endothelial BPAECs were relatively unscathed. (c) xz visualisation of the untreated co-culture reflects the integral layers shown in the xy z-stack images of (a). (d) xz visualisation of the BCG infected co-culture, including regions of catastrophic damage to the epithelial BATII layer (white brackets), whilst CD31 staining is unaffected, suggesting that detrimental effects of BCG infection are limited to the epithelial component of the model. Images acquired using a 20 × objective on a Nikon Ti confocal microscope.
