Fig. 2.

Transduction of carcinoembryonic antigen (CEA) into dendritic cells (DCs). (a) Flow cytometric analysis of CEA in DCs transduced with CEA or Tat-CEA. DCs incubated with either purified CEA or Tat-CEA protein for 10 min (left panel) or 30 min (right panel) were stained with anti-mouse CD11c-phycoerythrin (PE), followed by intracellular staining of CEA with anti-human CEA-fluorescein isothiocyanate (FITC) for fluorescence activated cell sorter (FACS) analysis. CD11c^pos cells were gated for the histograms. Percentage (%) of CD11c^posCEA^pos cells were analysed using gates (b + c) indicated. Filled histograms indicate isotype controls. (b) Confocal microscopic analysis of DCs transduced with recombinant CEA or Tat-CEA. DCs incubated with recombinant antigens were stained with anti-human CEA-FITC (green), anti-CD11c-Alexa Fluor 647 (blue) and Lo-Pro3 (red, to stain nucleus). Autofluorescence of cells for FITC channel (anti-CEA) was confirmed by staining DC (medium) with anti-CEA-FITC (boxed figure in CEA-30 min-merge panel).