Transcriptome analysis of high humidity-regulated genes in Arabidopsis thaliana

Arabidopsis Col0 were grown in pots for 12 days before the transfer to a humidity-controlled chamber set at relative humidity 30% for additional 2 days. High humidity treatments were performed for 0, 10-min and 30 min in a chamber with high humidity (higher than 90%). Triplicate experiments were performed for each time point.Total RNA was isolated using the Qiagen RNeasy Plant Mini Kit. The RNA was then treated with TURBO DNase (Thermo Fisher Scientific). The quality of DNA-free RNA was assessed using Bioanalyzer 2100 (Agilent, Palo Alto, CA, USA). PolyA selection was conducted using Dynabeads mRNA purification Kit (Thermo Fisher Scientific, Waltham, MA, USA) to isolate mRNA, followed by shearing to approximately 300-base fragments in size using Covaris S2 Ultrasonicator (Covaris, Woburm, MA, USA). RNA-seq libraries were constructed using MGIEasy RNA directional library prep kit (Complete Genomics Inc, San Jose, CA, USA). For RNA-seq sequencing, samples were run multiplexed on the MGI DNBSEQ-G400RS FCL flowcell at 100-bp PE length.