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. 2016 Dec 8;36(3):361–373. doi: 10.15252/embj.201592426

Figure EV4. Analysis of DNA covalently linked with Top1 by immuno‐pull‐down.

Figure EV4

  1. Immunoblot of yeast cell lysates of different strains probing for the presence of 5×FLAG‐tagged Top1 before (−) and after (+) immuno‐pull‐down with anti‐FLAG antibodies.
  2. The DNA covalently linked with Top1 purified by immuno‐pull‐down (after proteinase K digestion) was further treated with 3 μM recombinant TDP1 overnight or mock‐treated (Huang et al, 2013). TDP1‐treated DNA showed increased gel mobility. The control DNA was purified from the same sample after the immuno‐pull‐down procedure and TDP1 treatment showed no effect on the control DNA.
  3. A representative gel image of mock‐treated or S1‐treated pulled‐down DNA from either wild‐type (WT) or pol2‐M644G rnh201Δ strains.