Detecting Activated MAPKs and ROS in Arabidopsis

For the detection of activated MAPKs, immunoblot analyses using the anti-phospho p44/42 MAP kinase antibody (Cell Signaling Technology) were performed as described (Zhang et al., 2013 (link)). The detection of ROS in Arabidopsis leaf pieces was performed as described (Wan et al., 2019 (link)). Callose deposition in Arabidopsis leaves was stained with aniline blue 24 h after MAMP infiltration as described (Gomez-Gomez et al., 1999 (link)). Digital pictures of stained leaves were taken under UV-light with the microscope and callose depositions were quantified by counting light, fluorescent pixels with the Adobe Photoshop CS “select range” tool. Per treatment at least 10 pictures of at least 5 different leaves were analyzed.
For transcript profiling, RNA isolation and RT-qPCR analysis of plant material were performed as described previously (Zhang et al., 2013 (link)). The sequences of the primers used for PCR amplifications are listed in Supplementary Table S1.