Quantitative RT-PCR Analysis of Gene Expression

RNA was extracted with the RNeasy RNA isolation kit (Qiagen, cat# 74106). Reverse transcriptase reactions were performed using a SuperScript First‐strand Synthesis System (Invitrogen, cat# 12574026). Real‐time PCR reactions were performed on an ABI Prism 7500 system with the following primers: human CXCL14, 5′‐CGCTACAGCGACGTGAAGAA‐3′ and 5′‐GTTCCAGGCGTTGTACCAC‐3′; human BRG1, 5′‐TCATGTTGGCGAGCTATTTCC‐3′ and 5′‐GGTTCCGAAGTCTCAACGATG‐3′; mouse Cxcl14, 5′‐GAAGATGGTTATCGTCACCACC‐3′ and 5′‐CGTTCCAGGCATTGTACCACT‐3′; mouse Brg1, 5′‐CAAAGACAAGCATATCCTAGCCA‐3′ and 5′‐CACGTAGTGTGTGTTAAGGACC‐3′; mouse Il1b, 5′‐GCAACTGTTCCTGAACTCAACT‐3′ and 5′‐ATCTTTTGGGGTCCGTCAACT‐3′; mouse Il6, 5′‐TGGGGCTCTTCAAAAGCTCC‐3′ and 5′‐AGGAACTATCACCGGATCTTCAA‐3′; mouse Tnfa, 5′‐CTGGATGTCAATCAACAATGGGA‐3′ and 5′‐ACTAGGGTGTGAGTGTTTTCTGT‐3′; mouse Nos2, 5′‐GTTCTCAGCCCAACAATACAAGA‐3′ and 5′‐GTGGACGGGTCGATGTCAC‐3′; mouse Mcp1, 5′‐AAAACACGGGACGAGAAACCC‐3′ and 5′‐ACGGGAACCTTTATTAACCCCT‐3′; mouse Fasn, 5′‐GGAGGTGGTGATAGCCGGTAT‐3′ and 5′‐TGGGTAATCCATAGAGCCCAG‐3′; mouse Scd1, 5′‐ACTGTGGAGACGTGTTCTGGA‐3′ and 5′‐ACGGGTGTCTGGTAGACCTC‐3′; mouse Acc1, 5′‐GCGGCTACAGGGACTATACTG‐3′ and 5′‐CGGAAGTAAGAGCTACTAGCGG‐3′; mouse Srebp1, 5′‐TGACCCGGCTATTCCGTGA‐3′ and 5′‐CTGGGCTGAGCAATACAGTTC‐3′. Ct values of target genes were normalized to the Ct values of house keeping control gene (18 s, 5′‐CGCGGTTCTATTTTGTTGGT‐3′ and 5′‐TCGTCTTCGAAACTCCGACT‐3′ for both human and mouse genes) using the ΔΔCt method and expressed as relative mRNA expression levels compared to the control group which is arbitrarily set as 1.