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. 2016 Jun 2;291(30):15714–15726. doi: 10.1074/jbc.M116.730366

FIGURE 4.

FIGURE 4.

FeBABE-mediated hydroxyl radical cleavage of Mot1. A, schematic showing the positions of FeBABE molecules on the DNA probes. Lollipops represent FeBABE molecules coupled to phosphorothioates on the DNA 7 and 9 (4Fe), 20 and 22 (6Fe and 6GapFe), and 1 and 3 (10Fe) base pairs upstream of the TATA sequence. B, Western blot showing FeBABE-mediated cleavage of Mot1 in reactions containing streptavidin beads, phosphorothioate-conjugated biotinylated DNA, FeBABE, sodium ascorbate, H202, TBP, and Mot1. ADP-AlF4 was added as indicated. The numbers on the right represent approximate molecular masses of the Mot1 cleavage products in kDa. A unique band at ∼34 kDa was generated in reactions performed with probe 6Fe and ADP-AlF4 (lane 2) as shown by the cyan arrow. C, Western blot as in B but comparing the 6Fe and 6GapFe probes. The numbers on the right represent the approximate molecular masses of the Mot1 cleavage products in kDa. A unique band at ∼34 kDa appeared in reactions that contained ADP-AlF4 (lanes 2 and 4) as shown by the cyan arrow.