HDAC10 deletion in Treg

HDAC10–/– (B6N(Cg)-Hdac10tm1.1(KOMP)Mbp/J) and wild type (WT) CD4+CD25+ Treg were isolated and RNA procured. Single strand cDNA was generated, fragmented and labeled, and hybridized to Affymetrix GeneChip Mouse Gene 2.0 ST Arrays for microarray analysis. WT or HDAC10–/– Treg cell RNA was isolated using RNeasy kits (QIAGEN). Total RNA was assessed by UV absorbance and all had 260/280 nm ratios above 1.7. Additionally, samples were run on the Agilent RNA 6000 Nano chip and all had RINs in the 8.0 range. Microarray experiments were performed using whole-mouse-genome oligoarrays (GeneChip™ Mouse Gene 2.0 ST Array), and array data were analyzed using Transcriptome Analysis Console 4.0 software (ThermoFisher Scientific). Array data were subjected to robust multiarray average normalization. To assess differential gene expression, fold changes of up- and downregulated genes were calculated, and significance assessed via one-way ANOVA. Data with a false discovery rate <0.1 and >1.5-fold differential expression were included in the analysis.