Dexamethasone (DEX) induction, total RNA extraction and qRT-PCR

In order to induce the function of GR-RBE, DEX (10 μM dexamethasone, 0.1% ethanol, 0.015% silwet) or mock (0.1% ethanol, 0.015% silwet) solution was applied to 35S:GR-RBE young floral buds. After a 4 hour treatment, floral tissues were harvested and snap-frozen with liquid nitrogen. RNA was extracted with Trizol (Life Technologies), purified using TURBO DNA-free Kit (Life Technologies), and reverse transcribed with Multiscribe reverse transcriptase (Life Technologies) following the manufacturer’s protocols. qRT-PCR was carried out using the Taqman gene expression assay (Life Technologies). Gene expression levels were calculated from three biological replicates using the 2^–ΔΔC _T method (Livak and Schmittgen, 2001). The relative RNA levels were normalized to the value of ACTIN 2 (ACT2).