Fig. 4: Deletion of Arntl increased Fgf21 expression levels indirectly via REV-ERBα in adipocytes. | npj Biological Timing and Sleep

Fig. 4: Deletion of Arntl increased Fgf21 expression levels indirectly via REV-ERBα in adipocytes.

From: Deletion of Arntl, a component of the molecular clock, in adipocytes leads to cellular hypertrophy by increasing insulin sensitivity via FGF21

Fig. 4

a Expression level of genes related to insulin sensitivity in the eWAT of Arntlflox/flox and AAKO mice at ZT10 (n = 5). b Circadian gene expression levels in the eWAT of Arntlflox/flox and AAKO mice (n = 3–5). c Representative western blot of FGF21 in the eWAT. β-ACTIN was used as a loading control. d Serum FGF21 levels in Arntlflox/flox and AAKO mice (n = 3–5). e Circadian gene expression levels of Fgf21 in the liver of Arntl flox/flox and AAKO mice (n = 3). f Gene expression levels of Nr1d1, Arntl, Fgf21, Nfil3, and Rora in negative control (NC) siRNA- or Nr1d1 specific siRNA-transfected 3T3-L1 adipocytes. g Gene expression levels of Fgf21 in NC siRNA- or Nr1d1 specific siRNA-and pcDNA3.1 empty or Arntl expression vector transfected 3T3-L1 adipocytes. h Top, schematic representation of the mouse Fgf21 promoter region. Bottom, chromatin immunoprecipitation (ChIP)–quantitative polymerase chain reaction (qPCR) analysis of the interaction between REV-ERBα and the retinoic-acid-receptor-related orphan-receptor-responsive element (RORE) in the mouse Fgf21 promoter region in the eWAT at ZT10 and ZT22 (bottom. n = 3). Non-immune IgG sample was used to normalize the qPCR results in each ChIP sample. The Arntl RORE region and Fgf21 non-RORE region were used as the positive and negative control, respectively. Data are presented as the mean ± SD. Data in (a, f) were compared using an unpaired Student’s t-test with Welch’s correction. *P < 0.05, **P < 0.01 relative to Arntlflox/flox mice (a) or NC siRNA-transfected 3T3-L1 adipocytes (f). Data in panels were compared using two-way ANOVA with Bonferroni’s (b,d,e) or Tukey’s (g,h) post-hoc test. *P < 0.05, **P < 0.01, ***P < 0.001 relative to Arntlflox/flox mice at the same timepoint (b,d,e,h) or NC siRNA- and pcDNA3.1 empty vector transfected 3T3-L1 adipocytes (g), ##P < 0.01, ###P < 0.001 relative to ZT10 for the same genotype (h). Rhythmicity was calculated using CircWave software. R, rhythmic expression (P < 0.05); N non-rhythmic expression (P > 0.05).

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