Fig. 9. Intracellular targeting of CMG-1-GFP and CMG-2-GFP fusion proteins. CMG-1-GFP and CMG-2-GFP fusion protein vectors were created by preparing recombinant adenoviral vectors encoding the fusion proteins. A control GFP-only virus was also constructed. Recombinant adenoviruses were infected into 293 cells (a-d) or endothelial cells (e-g). In a-c, individual cells were photographed 24 hours after infection with CMG-1-GFP virus. Arrowheads represent fluorescent vesicular structures. In d, individual cells were photographed 24 hours after infection with GFP only virus. Bar, 20 µm (a-d). In e-g, endothelial cells were infected with CMG-2-GFP virus for 24 hours and were fixed and permeabilized for immunofluorescence staining. Panel e shows cells expressing the CMG-2-GFP protein. Panel f shows cells stained with anti-Hsp47 antibodies followed by staining with rhodamine-conjugated rabbit anti-mouse antibodies. No background staining was observed by the addition of the secondary antibodies alone. In g, a double exposure of the same field photographed in e and f is shown, illustrating the colocalization of CMG-2-GFP and Hsp47. Bar, 20 µm (e-g).