Fig. 7: PIP1s interact with SERK3 (BAK1), and PlPeL1/PlPeL1-like do not disrupt the LcPIP1-LcSERK3 interaction.
a LcPIP1-Flag was co-expressed with NbSERK3A-RFP, NbSERK3B-RFP, NbBAK1a-RFP, NbBAK1b-RFP, or RFP in N. benthamiana leaves. Protein complexes were immunoprecipitated with RFP-Trap-M beads. Co-precipitation was detected by western blot. b LcPIP1 interacted with LcSERK3 and NbSERK3A in vitro. GST-LcSERK3-, GST-NbSERK3A-, or GST- bound beads were incubated with bacteria lysate containing His-LcPIP1. Co-precipitation was detected by western blot. c PIP1s interacted with SERK3s in planta. SERK3 (BAK1) was co-expressed with PIP1-RFP or RFP in N. benthamiana leaves. Protein complexes were immunoprecipitated with RFP-Trap-M beads and detected by western blot. d Schematic representation of LcSERK3, featuring its extracellular leucine-rich repeat (LRR) domain in green, and intracellular kinase domain in blue. Transmembrane (TM) segment in gray and signal peptide (SP) in purple. e LcPIP1-HA was co-expressed with LcSERK3-RFP, LcSERK3ECD-RFP, LcSERK3CD-RFP, or RFP in N. benthamiana leaves. Protein complexes were immunoprecipitated with RFP-Trap-M beads. Co-precipitation was detected by western blot. f, g PlPeL1/PlPeL1-like did not disrupt the LcPIP1-LcSERK3 interaction. HA-tagged PlPeL1 or PlPeL1-like was co-expressed with Flag-tagged LcPIP1 and RFP-tagged LcSERK3 in N. benthamiana. Protein complexes were immunoprecipitated with RFP-Trap-M beads. Co-precipitation was detected by western blot. Red asterisks indicated protein bands of the correct size. Ponceau S staining of Rubisco was used to indicate loading quantity of protein samples. These experiments were repeated three times with similar results.
