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Fig. 3. RanGTP and importins {alpha} and ß colocalize to spindle poles. (A) The AR-12 antibody was used in unfixed cells to label native RanGTP, followed by FITC- (a) or rhodamine- (b) conjugated secondary antibody. Importin ß was revealed using Texas-Red- (a) or FITC- (b) conjugated secondary antibody. Scale bar, 10 µm. (B) Importin {alpha} in PFA-fixed metaphase (left) and anaphase (middle) cells; after Triton X-100 extraction (right), a pole-associated fraction is evident at spindle poles. Scale bars, 10 µm.