ChIP assays were performed as previously described (Bowler et al., 2004 (link); Perrella et al., 2018 (link); van der Woude et al., 2019 (link)). DNA was sheared using an ultrasonic bath (FALC instruments LABSONIC LBS2.10), cooled at 4°C using the maximum kHz frequency of 20 cycles 45 s ON, 15 s OFF. Anti-6XHis tag (Abcam ab9108) was used to IP the chromatin according to the manufacturer's instructions. ChIP-qPCR was performed using the following cycles: 95°C × 2 min, 95°C × 3 s, 59.5°C × 30 s for 50 cycles, 95°C × 1 min, and 60°C × 30 s to estimate the melting curve. Oligonucleotides for YUCCA8 and PIF4 DNA genomic regions were designed based on Lee et al. (2014) (link) and Zhu et al. (2016) (link). Relative ChIP DNA abundance was calculated as previously described (Kaiserli et al., 2015 (link)). In detail, MED25 enrichment over loci was determined by normalizing immuno-precipitated DNA against genomic DNA for the indicated regions and temperature conditions and indicated as a percentage of nuclear DNA (%Input). DNA was also immunoprecipitated from Col-0 wild-type plants and used as a negative control for ChIP-qPCR amplifications.