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Fig. 4. Bet3 is distributed in the cytosol. (A) NRK cells were fixed, permeabilized and double labeled with Bet3 antibodies (a and d) and a monoclonal antibody against ß-COP (b and e). The cytosolic labeling of Bet3 (d) but not ß-COP (e) was abolished by prior incubation of the antibodies with Bet3-His6. The merged images are shown in c and f. Bar, 10 µm. (B) Double labeling of Bet3 antibodies (a and d) and GS28 antibodies (b and e). NRK cells were permeabilized with saponin after (a-c) or before (d-f) fixation with paraformaldehyde. The merged images are shown in c and f. Bar, 10 µm. (C) HeLa cells were transiently transfected to express GFP-Bet3, which displayed cytosolic distribution in live cells expressing low-to-medial levels of GFP-Bet3. Some nuclear accumulation was detected in cells expressing very high levels (marked by white arrow).
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