Supplementary Figure 1: Hes1 and Hey1 suppress inflammatory gene expression in a selective manner.

(a) Quantitative RT-PCR (qPCR) analysis of mRNA expression of Hes1 and Hey1 in BMDMs from Hey1^+/+Hes1^fl/fl (WT) and Hey1^−/−Hes1^fl/flMx1-Cre (DKO) littermates. (b,c) Heat maps of gene expression profiles of LPS-induced genes (b) and LPS-suppressed gene (c) in WT and DKO BMDMs treated ± LPS (2 ng/ml) for 3 h (cut off=2 fold). Microarray analysis was performed as described in Methods. Hes1-regulated genes in both positive and negative directions are shown in enlarged panels on the right. An individual gene may be represented more than once due to presence of multiple probe sets in microarrays. 3 determinants of the Cxcl1 gene are marked with arrows (b, enlarged). The regulated gene number corresponds to unique genes but may include ESTs with uncharacterized functions shown without identifiers. Genes were ranked according to the order of numeric identifiers of microarray probes. (d) qPCR analysis of Cxcl1 mRNA expression in WT and DKO BMDMs stimulated with 10 ng/ml of Pam3Cys (TLR2 agonist) or 1 μg/ml of R848 (TLR7/8 agonist) for 3 h. Cumulative data from four independent experiments (a) or representative data from two independent experiments (d) are shown as mean and s.d.. Error bars are technical triplicate determinants in d. ***P<0.001 (two-tailed Student’s paired t-test). N.D., not detected.