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. 2010 Jul 20;107(31):13960–13965. doi: 10.1073/pnas.1002828107

Fig. 4.

Fig. 4.

SNC1 interacts with TPR1. (A) Coimmunoprecipitation of snc1-HA with TPR1-GFP in protein extracts of TPR1-GFP and snc1-HA double-tagged transgenic plants. Total protein extracts were subjected to immunoprecipitation with anti-GFP magnetic beads as previously described (32). Crude lysates (Input) and immunoprecipitated proteins (elution) were detected with anti-GFP or anti-HA antibodies. (B) In vitro analysis of the interaction between TPR1 and the TIR domain (amino acid 1–182) of SNC1. Crude lysates of E. coli expressing GST-tagged TIR domain of SNC1 (GST-TIR) or GST were mixed with crude lysates of E. coli expressing TPR1 before GST pull-downs. Aliquots of the mixtures (input) and proteins pulled down by GST were subjected to anti-TPR1 immunoblot analysis. GST-TIR and GST were detected by Ponceau staining. The polyclonal anti-TPR1 antibody was generated in rabbit using an N-terminal fragment (amino acid 1–356) of TPR1 expressed in E. coli.