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. 2015 Aug 14;27(8):2301–2313. doi: 10.1105/tpc.15.00444

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Figure 2. — LUH Interacts with PIF1.

(A) GST pull-down assay showing the interaction between recombinant His-tagged LUH (LUH-His) and GST-tagged PIF1 (GST-PIF1). Resin-bound GST or GST-PIF1 proteins were mixed with LUH-His and precipitated. Precipitated proteins were then analyzed by immunoblot using an anti-His antibody. The lower panel shows precipitated GST and GST-PIF1 proteins stained with Ponceau S.

(B) In vivo binding assay between FLAG-tagged LUH (LUH-FLAG) and Myc-tagged PIF1 (PIF1-Myc). Seven-day-old, light-grown, stable, transgenic lines expressing Myc-tagged GFP (GFP-Myc) or PIF1-Myc were treated to transiently express LUH-FLAG by Agrobacterium infiltration. Myc-tagged proteins were immunoprecipitated with an anti-Myc antibody and coimmunoprecipitated LUH-FLAG was detected with an anti-FLAG antibody. IN, input samples; IP, immunoprecipitated samples. Both in vitro and in vivo binding assays were performed at least three times, and representative data are shown.