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Figure 3


Fig. 3. Crosslinking of R4op. (A) Crosslinking of cytosolic R4op in the presence or absence of nucleotides. After termination of R4op translation in RRL, nucleotides were removed and either no nucleotides or 2 mM ATP or ADP added. After the crosslinking by either DSS (D) or BMH (B), R4op was immunoprecipitated and characterised by SDS-PAGE and autoradiography. (B) Crosslinking of R4op in the presence or absence of ribosomes. After translation, ribosomes were removed by ultracentrifugation. A total fraction (total), the resuspended pellet (pellet) and the supernatants (Sn) were crosslinked by BMH or left uncrossliked (–). (C) Crosslinking of R4op in the presence of Triton X-100. After synthesis of R4op in the RRL, the samples were adjusted to the indicated amount of Triton X-100 and then BMH was added. (D) Crosslinking of R4op in the absence or presence of RMs. After synthesis of R4op, BMH was added either immediately or after the additional incubation with RMs. g-R4op, glycosylated R4op. R4op x p40, crosslinked product of R4op and a 40-kDa protein.





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