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Figure 8


Fig. 8. Co-immunostaining of G{alpha}12 and G{alpha}o in cells exposed to a UTP gradient. Human 1321N1 cells expressing wild-type P2Y2Rs were plated on chambered coverslips and incubated overnight in 300 µl serum-free medium. UTP (1 µl of a 100 mM solution) was added at the bottom left and after 30 minutes, cells were washed, fixed, permeabilized and stained with rabbit anti-G{alpha}12 and mouse anti-G{alpha}o antibodies. Texas-Red-conjugated anti-rabbit IgG and Alexa Fluor 488-conjugated anti-mouse IgG were then used to localize G{alpha}12 and G{alpha}o, respectively. Images representative of three experiments are shown. Arrows indicate membrane protrusions indicative of lamellipodia formation. Bars, 50 µm (left column); 20 µm (enlarged images in right column).





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