Figure 6. APX2 and HSFA3 are two direct binding targets of BES1.
(A). Activation of BES1 against promoters of HS-associated memory genes was tested in Y1H experiment. Growth of yeast transformants carrying the HIS3 reporter gene under the control of each promoter region was examined in the medium lacking leucine (L), Tryptophane (W) and histidine (H) with or without 3-AT.
(B). Electrophoretic mobility shift assay. MBP-BES1 protein binds specifically to the BES1 binding site within the APX2 promoter. 2+ represents twice the amount of protein, m represents the probe mutated BES1 binding site.
(C). Electrophoretic mobility shift assay. MBP-BES1 protein binds specifically to the BES1 binding site within the HSFA3 promoter. 2+ represents twice the amount of protein, m represents the probe mutated BES1 binding site.
(D). ChIP-qPCR assays indicate enrichment of APX2 andHSFA3 promoter in the immunoprecipitation products of BES1 protein. The ChIP assays were performed with chromatin prepared from35S::GFP and 35S::BES1-GFP plants harvest at 12 h after heat priming, using an anti-GFP antibody. The precipitated DNA was analyzed by qPCR using the primer pairs of HS-associated memory genes. P-ACTIN2 was used as the negative control. Values are means ± SD of three independent biological replicates.