FIG. 5.

Biochemical requirements for recruitment of p73 onto the ER membrane. Sixteen hours after infection with the BA71v strain of ASF virus, Vero cells were pulse-labeled with [³⁵S]methionine and cysteine for 5 min at 37°C and then chased for increasing times under the conditions described in the legends to Fig. 3 and 4. Cells taken at each time point were homogenized, and postnuclear-membrane and cytosolic fractions were lysed in immunoprecipitation buffer. The levels of p73 present were estimated by immunoprecipitation followed by SDS-PAGE and autoradiography. (A) The distribution of p73 between membrane (M) and soluble (S) fractions was determined for control cells by immunoprecipitation followed by SDS-PAGE and autoradiography. (B) The levels of p73 present in the membrane fraction were estimated by densitometry, and the percentage of the total p73 recovered at each time point is presented for control cells (■) and cells incubated with 50 mM 2-deoxy-d-glucose and 1 mM sodium cyanide (○) or 1 μM A23187 and 3 mM EDTA (●).