Anti bik1

Four-week-old plant leaves were infiltrated with sterile water (mock) or different Pst strains at OD₆₀₀=0.02, and samples were collected at 0.5, 3, 6, 8h after infiltration. Three to four leaves from different plants were collected as one sample. Protein was extracted using Plasma Membrane Protein Isolation Kit (Invent) according to the manufacturer’s protocol. Concentration of the cytosolic protein was determined with Bradford protein assay kit (Bio-Rad). An equal amount of protein was loaded onto SDS-PAGE gel for western blot. BAK1 and RBOHD are detected in the immunoblot of total membrane fraction and other proteins are detected in the immunoblot of total protein extracts. Different PTI components were detected by following antibodies with indicated dilution: anti-RBOHD (Agrisera), 1:1000; anti-BAK1 (Agrisera), 1:5000; anti-BIK1 (Agrisera), 1:3000; anti-MPK3 (Sigma-Aldrich), 1:2500; anti-MPK6 (Sigma-Aldrich), 1:5000. Goat Anti-Rabbit lgG HRP (Abmart; 1:5000) was used as secondary antibody. The protein image was taken using the Tanon-5200 imaging system (Tanon).