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Figure 9


Fig. 9. Autoradiogram of intestinal ER under conditions favoring PCTV budding identifies a 9-kDa phosphorylated protein. Intestinal ER (500 µg) was incubated for 30 minutes at 37°C with native intestinal cytosol (1 mg) and an ATP-regenerating system including 20 µCi [{gamma}-32P]ATP. The reaction was stopped by placing the incubation tube on ice and adding cold 10 mM HEPES buffer (pH 7.2). 20 µl were solubilized in Laemmeli buffer and the proteins separated by 15% SDS-PAGE. The gel was dried and autoradiography performed at –70°C for 10 days. (A) Complete reaction, incubated at 37°C in triplicate. (B) Complete reaction incubated at 4°C. (C) PCTV-budding reaction without cytosol incubated at 37°C. (D) PCTV-budding reaction without ER incubated at 37°C.





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