Genome-wide RNA-sequencing of mouse islets 48 hour after transduction with adenoviruses expressing either GFP (control), or constitutively active forms of the transcription factors, Nfatc1 or Nfatc2.

Genetic variation at ~160 gene loci is associated with type 2 diabetes (T2D). Using an F2 mouse intercross segregating for T2D, we searched for a driver of GWAS gene expression and found that ~40% of the GWAS genes are regulated in trans by a locus on chromosome 2 in islets. We identified Nfatc2 as a candidate driver of GWAS gene expression. Overexpression of Nfatc2 induced ß-cell proliferation in mouse and human islets. We show that many T2D GWAS genes are responsive to NFAT and thus may act coordinately as intermediate traits to promote diabetes. Overall design: On each of 5 separate days ~1,200 islets were isolated and pooled from 6 B6 mice, and then separated into 3 portions for adenoviral transduction to overexpress GFP (control), or constitutively active (ca) forms of mouse Nfatc1 or Nfatc2. 48 hr later, whole islet RNA was isolated using RNeasy purification columns (Qiagen), quantified (Nanodrop) and integrity verified (Agilent) prior to sequencing. The 15 separate RNA samples (N = 5 each for GFP, ca-Nfatc1 and ca-Nfatc2) were bar-coded and randomized for multiplexing across three lanes of an Illumina HiSeq 2000, which yielded ~24M total RNA-sequencing reads/sample.