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. 2011 Jan 27;6(1):e14605. doi: 10.1371/journal.pone.0014605

Figure 6. Expression of Rcan2 mRNA in the hypothalamus and its involvement in the hyperphagic responses.

Figure 6

(A) Expression of hypothalamic Rcan2-1 and Rcan2-3 mRNAs in the ad lib fed state. Total RNA was isolated from hypothalami of 6-week-old wild-type mice or 9-month-old wild-type mice that was ad lib fed with the normal chow diet (NCD), or from hypothalami of wild-type mice that had been fed with the high-fat diet (HFD) for 5 months. The RNA was used for quantitative real-time PCR analysis using primers specific for Rcan2-1, Rcan2-3 and β–actin (internal standard). mRNA expression levels are expressed relative to those of 6-week-old mice. NCD-6W indicates 6-week-old mice fed with the NCD; NCD-9 mon, 9-month-old mice fed with the NCD; HFD-5 mon, wild-type mice fed with the HFD for 5 months. n = 5 in each group (B) Expression of hypothalamic Rcan2-1 and Rcan2-3 mRNAs in the fasted state. Total RNA was isolated from hypothalami of wild-type male that were allowed to feed ad lib or were fasted for 24 h. The RNA was used for quantitative real-time PCR analysis using primers specific for Rcan2-1, Rcan2-3 and β–actin (internal standard). mRNA expression levels are expressed relative to those of ad lib fed wild-type mice. Data were expressed as the mean of three independent experiments. Thirty-one ad lib fed wild-type mice and 30 fasted wild-type mice were used in this analysis. Red columns indicate the mRNA expression levels in the ad lib fed state; blue columns, the mRNA expression levels in the fasted state. (C) Cumulative food intake in response to 24-h fasting. Six wild-type males (28.90±0.67 g) and six Rcan2 −/− males (24.78±0.81 g) were analyzed. (D) Body weight reduction after 24-h fasting and 24-h refeeding. Six males of each genotype were analyzed. All values are given as mean ± s.e.m. *: P<0.05; **: P = 0.01; ***: P<0.0001. Filled circles/columns indicate Rcan2 +/+ mice; open circles/columns, Rcan2 −/− mice.