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. 2017 Nov 15;6:e27434. doi: 10.7554/eLife.27434

Figure 3. Pulldown of core BBSome with immobilized Arl6 and Smoothened domain R546-L665.

Figure 3.

Purified core BBSome samples were pulled down by a 7.5 fold molar excess of Arl6•GDP, Arl6•GppNHp or Smo(R546-L665) immobilized to Streptactin-coupled Sepharose beads. While no interaction of the BBSome with Arl6•GDP was detectable, Arl6•GppNHP captured most of the BBSome sample. Smo(R546-L665) bound weaker, but still captured >2% of the BBSome sample. This was approximated by a comparison of the 1:50 diluted Arl6•GppNHp elution with the undiluted Smo(R546-L665) elution in a Flag-blot against Flag-BBS8 from the BBSome core complex.