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. 2012 Jul 9;109(31):12662–12667. doi: 10.1073/pnas.1209965109

Fig. 1.

Fig. 1.

Lack of a functional circadian clock system by absence of the circadian oscillator component CRY constitutively upregulates expression of inflammatory cytokines. (A) Estimation of mRNA levels of indicated genes by quantitative real-time PCR (qRT–PCR) in the hypothalamus region of brain of WT and Cry1−/−;Cry2−/− mice are shown. Data are mean ± SEM (n = 4 mice per group); *P < 0.05. (B) Quantification of mRNA levels of indicated genes by qRT–PCR in WT and Cry1−/−;Cry2−/− fibroblasts are shown. Data are mean ± SD (n = 3). (C) WT, Per1−/−, Per2−/−, Bmal1−/−, Cry1−/−, Cry2−/−, and Cry1−/−;Cry2−/− fibroblasts were left untreated for 12 h and the supernatants were analyzed for IL-6 protein levels by ELISA. Data are mean ± SD (n = 3). (D) BMDM from WT and Cry1−/−;Cry2−/− mice were analyzed for mRNA levels of indicated genes by qRT–PCR. Data are mean ± SD (n = 3). (E) WT and Cry1−/−;Cry2−/− BMDM cells were either untreated or treated with LPS (1 μg/mL for TNF-α and 10 ng/mL for IL-6 analysis) for 12 h and the supernatants were estimated for TNF-α or IL-6 protein levels by ELISA are shown. Data are mean ± SD (n = 4). Relative mRNA expression levels after normalization to actin are shown in A, B, and D.