Fig. 2. Expression of integrin Vß3 induced by RANKL-ODF treatment. (A) Chicken peripheral blood-derived macrophages were cultured on glass coverslips for four days in the presence (right panels) or absence (left panels) of RANKL-ODF (see Materials and Methods). Cells were fixed, permeabilized and stained with the monoclonal antibody 23C6 (green) and rhodamine-phalloidin (red). Confocal images were acquired using identical channel settings for Vß3 detection and were equally processed in Adobe Photoshop to ensure comparable detection of Vß3 expression. Upper panels, 23C6-staining only; lower panels, 23C6 + rhodamine phalloidin staining. (B) Human osteoclast obtained after a nine day treatment of peripheral blood monocytes with RANKL-ODF. Staining was performed as in A and revealed by confocal microscopy. Upper panel, rhodamine phalloidin-staining (red); lower panel, 23C6- (green) + rhodamine phalloidin-staining. Note the nearly complete absence of integrin Vß3 on cells that were not treated with (A, left panels) or that did not visually respond to (B) the osteoclastogenic factor RANKL-ODF. Scale bar: 20 µm).