Fig. 2

CgMkk1-mediated phosphorylation and localisation of CgCrzA under CFW Stress. A In vivo phosphorylation of CgCrzA by CgMkk1 detected using Mn²⁺-Phos-tag and conventional SDS-PAGE gel, with detection facilitated by an anti-GFP antibody. Phosphatase represents alkaline phosphatase treatment; PI indicates phosphatase inhibitors. B In vitro phosphorylation analysis conducted using the FDIT method. Error bars represent standard deviation. Asterisks indicate a significant difference at P < 0.01. C Localization of CgCrzA in the ΔCgmkk1 mutant observed in response to CFW. Scale Bar = 10 μm. D LC-MS/MS analysis identified CgCrzA phosphorylation sites in the wild-type and ΔCgmkk1 mutant expressing CgCrzA. E In vivo phosphorylation of CgCrzA by CgMkk1 is abolished upon mutation of the Ser280 residue. Variants of CgCrzA expressed in ΔCgcrzA and their phosphorylation level in response to CFW were detected using Mn²⁺-Phos-tag (upper) and normal SDS-PAGE (lower) gel