Extended Data Fig. 4: Identification of phosphorylation site within the BSR1 by LC-MS/MS.
a, b, In vitro phosphorylation of BSR1 CC and LRR domains by NbWAKL20. The kinase-dead mutant NbWAKL20K375E served as negative control. The autoradiographic signals (left) and the CBB-stained gel used as a loading control (right) are shown. c, Silver-stained SDS-PAGE gel image of BSR1-9×Flag protein immunoprecipitated from N. benthamiana leaf tissues. The band indicated by the arrow was excised for LC-MS/MS analysis. d, CBB-stained SDS-PAGE gel image showing MBP-NB-ARC or MBP-NB-LRR proteins after treatment with NbWAKL20KD with or without the addition of ATP. The band indicated by the arrowheads were excised for LC-MS/MS analysis. e, Summary of the phosphorylation sites within the BSR1 identified by LC-MS/MS. f, In vitro phosphorylation of BSR1 NB-ARC ___domain and its variants by NbWAKL20. The autoradiographic signals (left) and the CBB-stained gel used as a loading control (right) are shown.
