For data validation we analyzed results of ChIP assays repor

For data validation, we analyzed results of ChIP assays reporting allele-specific binding of ZFP57 to ICRs/gDMRs [15]. Our approach localized the likely peak-positions of the canonical ICRs/gDMRs in the mouse genome (Table 3); for details see Ref. [10].
Data The dataset of this article provides information on T-DNA insertions in SALK_084889. Fig. 1 shows the T-DNA bands amplified within DREB2A (AT5G05410) and LOX1 (AT1G55020) genes of SALK_084889 as well as T-DNA insertion sites determined by Sanger sequencing. Fig. 2A–B show DREB2A oxygenase in both roots and shoots in normal conditions with or without drought treatment. Fig. 2C–D show LOX1 expression in roots and shoots in normal conditions. Figs. 3 and 4 show the comparison of survival rates between Col and SALK_084889 plants in drought and heat treatments. Table 1 shows the sequences of primers used in experiments for Figs. 1 and 2.
Experimental design, materials and methods Arabidopsis seeds of wild tyoe (Col) and SALK_084889 were obtained from Arabidopsis Biological Resource Center. SALK_084889 was reported as a T-DNA insertion line of DREB2A (AT5G05410) gene [1]. For identification of T-DNA insertion sites, Col and SALK_084889 seeds were grown in soil at 22°C under constant light condition. Eight randomly selected SALK_084889 seedlings were subjected to DNA-PCR to confirm all seeds are homozygous. For DREB2A, the gene specific primers DREB2A-F and DREB2A-R were used to amplify wild type allele band while LBb1.3 and DREB2A-R primers were used to amplify T-DNA band. For LOX1, the gene specific primers LOX1-F and LOX1-R were used to amplify wild type allele band while LBb1.3 and LOX1-R primers were used to amplify T-DNA band. The amplicons of T-DNA bands of DREB2A and LOX1 were purified by subjected to sanger sequencing and the alignment of sequences was performed using Vector NTI v10.0 program (InforMax Inc). For Expression analysis, Col and SALK_084889 seeds were grown in 0.5× MS media (2.2g of MS basal salts (Sigma), 5g of Sucrose, 0.5g of MES, and 1mL of 1,000× Gamborg vitamins (Sigma) per liter at pH 5.75) at 22°C under constant light condition. The 14 day old plants were harvested from MS plates and directly preserved in RNALater (Ambion) [2]. For drought treatment, harvested Arabidopsis plants were dehydrated on Whatman 3mm paper (Whatman) at 22°C and 60% humidity under dim light for 2h [3] prior to fixation in RNALater. For RNA extraction, root and shoot tissues were dissected in RNALater solution and then total RNA was isolated using the Qiagen RNAeasy kit (Qiagen). Quantitative realtime PCR was performed using Fast SYBR Green Master Mix (Thermo Fisher) and 2-ΔΔCt method was employed to calculate the relative expression levels of DREB2A and LOX1. Three replicates were used for each sample and UBQ11 (AT4G05050) gene was used as the internal control. For drought and heat tolerance determination of SALK_084889, Col and SALK_084889 seeds were grown in soil at 22°C under constant light condition. For drought test, the 10 day old plants were transferred to pre-dried soil and withhold water for 4 days prior to recovery by watering for 7days [4]. For heat test, the 7 day old plants were pre-incubated in 37°C for 1h and then treated at 49°C for 1h prior to recovery in 22°C for 2 days [5]. Survival rates and phenotypes of Col and SALK_084889 plants were recorded. Student׳s t-test was used for statistical analysis.
Acknowledgements The authors thank the support from members of the UF Space Plants Lab. This work was funded by NASA Space Life and Physical Sciences Grants NNX13AM46G and NNA04CC61G awarded to RJF and ALP managed through Kennedy Space Center.
Data These data provide detailed information regarding foxtail millet CNL R-genes [1]. Generated from sequence annotations, gene structure diagrams for each of the 242 genes were compiled (Fig. 1), along with InterProScan and Gene Ontology (GO) annotations (Table 1). Utilizing CNL R-gene genomic locations, chromosomal syntenic maps (Fig. 2) and R-gene density (Fig. 3) are displayed. Based on R-protein sequences, homologs were compiled (Table 2) and a maximum-likelihood phylogenetic tree (Fig. 4) displays the evolutionary relationships between Setaria italica, Arabidopsis thaliana, Sorghum bicolor, and Panicum virgatum accessions.