DRE2 associates with chromatin and some auxin responsive genes.
(A-B) Detection of DRE2 in cytoplasmic, nucleoplasmic and chromatin-associated fractions by Western blot. DRE2-HA fusion protein was detected using an anti-HA antibody. DRE2-GFP fusion protein was detected using an anti-GFP antibody. HSP90 and histone H3 were detected as markers for cytoplasmic proteins and chromatin proteins, respectively. AGO1 was detected as markers for proteins present in all the fractions. (C) Immunostaining results showing the nuclear distribution of DRE2. DRE2-HA in the pDRE2::DRE2-HA transgenic plants was stained with an anti-HA antibody and an Alexa Fluor 488-conjugated secondary antibody (green). DNA was stained with DAPI (blue). (D) ChIP-qPCR results showing the enrichment of DRE2-GFP at four auxin responsive genes. (E) Relative expression levels of the indicated genes in the indicated genotypes as determined by RT-qPCR. Data are presented as mean ± SD of four technical replicates. Asterisks indicate two-tailed Student’s t-test, *P < 0.05, **P < 0.01.