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. 2013 Feb 19;110(15):6205–6210. doi: 10.1073/pnas.1215543110

Fig. 6.

Fig. 6.

PEPRs mediate BIK1 phosphorylation in response to Pep1 and ET. (A) The PEPR1 KD phosphorylates BIK1 in vitro (four independent experiments). GST-BIK1K105E was incubated with PEPR1-KD-His in a kinase buffer containing 32P-γ-ATP, and protein phosphorylation was detected by autoradiography. CBB indicates loading of the protein. (B) Pep1-induces BIK1 phosphorylation in plants (three independent experiments). BIK1::BIK1-HA transgenic plants (WT or pepr1/pepr2 background) were sprayed with H2O (-) or 10 μM Pep1, and tissues were harvested 15 min later. BIK1-HA phosphorylation was detected as a band-shift in an anti-HA immunoblot. PPase, protein phosphatase. (C) Pep1 induces BIK1 phosphorylation in ein3/eil1 plants carrying the BIK1::BIK1-HA transgene (two independent experiments). (D) ACC induces BIK1 phosphorylation in a PEPR1/PEPR2-dependent manner. BIK1::BIK1-HA transgenic plants of pepr1/pepr2 and WT background were treated with ACC for 2 h, and BIK1 phosphorylation was detected as by anti-HA immunoblot (three independent experiments).