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. 2012 Aug 22;287(42):35768–35778. doi: 10.1074/jbc.M112.396911

FIGURE 6.

FIGURE 6.

Effect of 3-AP on the diferric cluster of β2 immunopurified from treated and untreated 55Fe-pulse-labeled K562 cells. A, rapid immunoprecipitation was performed with anti-β2 antibody (Abcam) conjugated to Protein G-Dyna beads (Invitrogen). a, immunoprecipitated human β2 band (*, monomer, 45 kDa) was confirmed with Western blotting using human β2-specific antibody (Ab57653). b, Mr ladders (Bio-Rad). c, PVDF membrane in a was stained with Coomassie Brilliant Blue. * indicates immunoprecipitated β monomer. d, control as in a except buffer replaced Ab57653 but Protein G-Dyna beads were kept. B, shown is the amount of radioactivity associated with isolated β2 subsequent to 30-min treatment of the prelabeled cells with 5 μm inhibitor. Shown are the results from 3-AP-untreated (a) and -treated cells (b). c, treatment was identical to a, except anti-β2 antibody was omitted but the beads were retained. d, treatment was a except 10 mm HU was added to the lysate. Error bars are S.D. over duplicate sets of experiments.