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YB-1 interacts directly with pure tubulin. Affinity chromatography analysis of YB-1-tubulin interaction. Tubulin (20 μg) was applied onto 50 μl YB-1- or casein-Sepharose (used as a control) in buffer containing 150 mM NaCl at 4°C and incubated for 10 min. After washing, bound proteins were eluted stepwise using buffer containing 150 mM, 300 mM, 600 mM and 1 M NaCl as described under Materials and Methods. Fractions were analyzed by 15% SDS PAGE followed by Coomassie blue staining.
