Fig. 3.
AF-ALN is internalized by osteoblast-like cells and co-localizes with RPTPµ. To visualize the uptake of ALN by osteoblastic cells and its co-localization with a phosphatase, ROS 17/2.8 cells were incubated for 30 min with 0.1 µM Alexa Fluor 488-ALN (green), then fixed and incubated overnight with a specific antibody that recognizes RPTPµ, followed by an appropriate dilution of secondary antibody labeled with Alexa Fluor-647 (red). The samples were analyzed by confocal microscopy (scale bar, 31.75 µm). Phase contrast image (A); AF-ALN (B); RPTPµ (C); merge image showing co-localization (arrows) of pixels in yellow (D). (Images A and B are reprinted with kind permission of Elsevier, Archives of Biochemistry and Biophysics, 518 (2012) 95–102, Lezcano, Bellido, Plotkin, Boland and Morelli).