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. 2011 Dec 2;6(12):e28394. doi: 10.1371/journal.pone.0028394

Figure 3. Mixl1 DNA binding activity is not required for its association with Brachyury.

Figure 3

(A) Mixl1 DNA binding mutants and their interaction with T. 293T cells were transfected with GST mT and HA mMixl1 wt or Mixl1 HD DNA binding mutant expression plasmids as indicated in the text. GST-fusion proteins were isolated from whole cell extracts using glutathione resin and the bound fraction was analysed by Western blot with an anti-HA antibody. Expression of each protein was confirmed with anti-GST and anti-HA antibodies. (B) The Mixl1 P126I (PI) mutant shows reduced dimerisation. 293T cells were transfected with GST mMixl1 and HA mMixl1 wt and DNA binding mutant expression plasmids as indicated. Whole cell extracts were prepared and reactions performed as in (A).