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. Author manuscript; available in PMC: 2015 Jan 15.
Published in final edited form as: Curr Biol. 2011 Mar 22;21(6):520–525. doi: 10.1016/j.cub.2011.02.029

Figure 1. AFB4 and AFB5 Interact with ASK1 and Interact with the Aux/IAAs in an Auxin-Dependent Manner Revealing their Role as Auxin Receptors Pull-down experiments were carried out using crude plant extracts prepared from [tir1-1] GVG-TIR1-myc, [afb5-5] AFB5-AFB5-myc, and [afb5-5] AFB5-AFB4-myc seedlings and recombinant GST-IAA3.

Figure 1

(A) TIR1-myc, AFB4-myc, and AFB5-myc were immunoprecipitated with anti-myc antibody coupled to agarose beads, and ASK1 was detected with an anti-ASK1 antibody. The lower band corresponds to ASK1 protein.

(B) Pull-down reactions were incubated for 45 min in the presence or absence of 50 μM indole-3-acetic acid (IAA). GST-IAA3 was immunoprecipitated with glutathione agarose beads, and AFB4-myc and AFB5-myc protein were detected with anti-c-myc-peroxidase antibody.