Global gene expression profile of wild type and Smad4-deficient Th17 cells cultured at 37°C and 39.5°C

Febrile temperature enhanced Th17 differentiation in Smad4 SUMOylation-dependent manner. Smad4-deficiency impaired the febrile temperature-enhanced Th17 differentiation. In order to understand how febrile temperature and Smad4 contribute to Th17 pathogenicity, we conducted transcriptional analysis of wild type and Smad4-deficient Th17 cells cultured at 37°C and 39.5°C (Day 3). Naive CD4+ T cells were sorted from wild type and Smad4-deficient mice and differentiated into Th17 cells at 37°C and 39.5°C. The total RNA was extracted with Trizol (Life Technologies) according to manufacturer’s instructions, and the RNA-seq library was constructed with NEXTflex RNA-seq kit (Bioo Scientific) and sequenced with BGI500 platform by BGI Genomics and successfully mapped to mouse genome. Reads counts were normalized based on RPKM/FPKM, fold changes were calculated for all possible comparisons.