Figure 6.

GRP78 colocalization studies. Epifluorescence imaging of probe 5. a) An image collected after incubation of U2OS cells with 50 μM probe 5 for 75 min. After treatment the cells were fixed and staining with 1:20 dilution of the anti-GRP78 mAb (3177S, Cell Signaling) and 80 μM Alexa647-conjugated anti-IAF mAb (XRI-TF35, Xenobe Research Institute). Localization of GRP78 (red) was completed by staining with a Cy3B-labeled secondary antibody and imaging with λ_ex 535–585 nm and λ_em 600–660 nm. The localization of IAF tag (green) was conducted by imaging with λ_ex 590–650 nm and λ_em 663–738 nm b) Expansion of the image (white box) in a). Bars denote 10 μm.