HeLa cells were grown on a glass coverslip and incubated for 1 hour at 37°C with unlabeled ADDomer at 20 μg/ml. Subsequent to methanol fixation, rabbit serum raised against human Ad3 penton base protein, diluted 1:1000 in PBS, was applied, followed by Cy5-labeled goat anti-rabbit antibody (Jackson ImmunoResearch), diluted 1:500 in PBS. Nuclei were stained by the DNA dye 4′,6-diamidino-2-phenylindole.

Monocytes and MDDCs from healthy anonymous donors were obtained from the National Blood Service [Etablissement Francais du Sang (EFS), La Tronche] after written consent following approval by the EFS. ADDomer (1 mg/ml) was labeled with Alexa Flour 488 dye (Invitrogen) following the manufacturer’s recommendations. Unbound dye was removed by extensive dialysis against PBS overnight. About 105 monocytes or MDDCs, respectively, were incubated for 1 hour at 37°C with 10 μg of labeled ADDomer. Cells were applied to glass coverslips and analyzed using differential interference contrast and green fluorescence channels in an Olympus IX81 inverted microscope.

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