Transcriptional Activation Analysis of CBF3

This experiment was performed as described previously (Qian et al., 2019 (link)). PCR amplified and cloned the ADF5 promoter sequence into pGWB235-LUC to construct the reporter vector. The full-length CBF3 coding sequence was amplified using PCR, and it was cloned into the pBIB-35s-GWR-flag vector as the effect vector. The reporter and effector vectors were co-transformed into N. benthamiana leaves for transcriptional activation analyses. A reporting vector alone was used as a negative control. Fluorescence of the luciferase and luciferin (Promega) reaction was obtained using a Lumazone CA1300B camera (Photometrics). The primers used to clone are listed in Supplementary Table 1.