TaMYB4 and TaMDHAR Localization in Tobacco

The CDS of TaMYB4 and TaMDHAR were constructed into the pCAMBIA2300-EGFP vector. Tobacco leaves (30 days old) were then injected with Agrobacterium tumefaciens GV3101 harboring pCAMBIA2300-EGFP, pCAMBIA2300-TaMYB4-EGFP or pCAMBIA2300-TaMDHAR-EGFP plasmids. After 72 h of injection, the leaves were harvested to observe the EGFP signal under a fluorescence microscope. Western blot analysis was performed to further analyze the subcellular localization results. The detailed experimental methods refer to those used by the authors in [43 (link)]. Anti-EGFP antibody (1:1000, Proteintech, Wuhan, China) was used as the primary antibody to probe EGFP. Horseradish peroxidase (HRP)-conjugated anti-rabbit antibody was chosen as the secondary antibody (1:2000, Proteintech).