Def1 associates with polyribosomes. (A) Polysome profiling. Cell lysate was fractionated on a 10%–50% sucrose gradient as described in the Materials and Methods section. The A254 recording of the gradient fractions is displayed on the y-axis. Def1 and Rpl3-s (large subunit protein) were detected by western blotting. The locations of the mRNP, 40S, 60S, 80S monosomes and polysomes are indicated. Two blots were used to analyze all the fractions, and the splice is indicated with a line. An extract from a def1Δ strain was analyzed to validate the antibody. (B) Migration of Def1 in the presence (red) or absence (black) of the polysome-disrupting agent EDTA. EDTA was added to the extract and gradient to disrupt polysomes.
