TRAIL localization in DENV-2-activated pDCs by 3D microscopy analysis.

<p>Freshly purified pDCs stimulated with DENV-2 pre-treated or not with chloroquine (Chloro), or mock infected. TRAIL expression was analyzed by flow cytometry or by a 3D microscope. (<b>A</b>) Membrane TRAIL flow cytometry profiles (left column) on pDC stimulated by different stimuli -mock, DENV-2 or DENV-2+Chloro overlaid by unstimulated (grey). Microscopic images from pDC cultured with the mock, DENV-2 or DENV-2+Chloro showing DAPI-colored nucleus, TRAIL staining (green) and overlay. (<b>B</b>) 3D interactive surface plots analysis of 3D microscopic image. Overlay of nucleus (blue), TRAIL (green) and phase contrast (grey) as seen in (<b>C</b>) for different stimuli: DENV-2-stimulated pDC (DENV-2) exhibits membrane TRAIL localization in contrast to DENV-2+Chloro or unstimulated, where TRAIL is detected only intracellularly. (<b>D</b>) Percentage of pDCs expressing intracellular TRAIL only (Intra) or on the membrane (Mb) is shown as percentage of total analyzed cells. Values were submitted to paired t test in which * p<0.05 and *** p<0.0005.