Figure 3.

βC1 Interacts with AS1 but Not AS2. (A) In vitro pull-down assays. Two micrograms of GST or GST fusion proteins were used to pull down 2 μg of MBP or MBP fusion proteins. (B) Competitive pull-down assays. Indicated protein amounts of βC1(ΔN8) or SUMO were mixed with 2 μg of MBP-AS1 and pulled down by 2 μg of GST-AS2. After being pulled down, Western blottings were performed using anti-MBP antibody to detect the associated proteins. Membranes staining with Coomassie Brilliant Blue were used to monitor input protein amounts. Asterisks indicate degradation products of MBP-AS1. (C) In vivo interaction of βC1 and AS1 in N. benthamiana. Agrobacterium cultures carrying 35S:Myc-AS1 and XVE:HA-βC1 were suspended in 50 μM β-estradiol and coinfiltrated into tobacco leaves. Transiently expressed Myc-AS1 and HA-βC1 were analyzed by coimmunoprecipitation. Crude extracts (Input) were used for immunoprecipitation (IP) with or without polyclonal anti-myc antibody and analyzed by Western blottings. Asterisk indicates the cross-reacting band.