![]() ![]() (F) Cosinor analysis of clock gene mRNA and protein in LD and DD. (E) Direct comparison of LD and DD conditions (7 days) of qPCR-determined expression of clock genes in the liver for Liver-RE mice. (D) qPCR-determined expression of clock genes in the liver for WT and KO mice, n=3–4. (C) Daily average caloric intake of mice subjected to metabolic cages under constant darkness. (B) Energy expenditure group averages over 2 days (top) and quantification of subjective light and dark periods (bottom). (A) Food intake for individual mice over a 2-day period in constant darkness. Characterization of Liver-RE mice under constant conditions. TSS = transcription start site.įigure S6. (H) BMAL1 occupancy at distinct groups of oscillating genes, for comparison of all groups and time points. (G) Proportion of published REV-ERBα binding sites (GSE number listed) associated with autonomously or non-autonomously oscillating genes. (D-F) Examples of peaks found only in WT, only in Liver-RE, or near core clock genes. (C) Pie chart of the genomic distribution of binding sites within promoter, genic or intergenic regions in WT or Liver-RE at ZT20. (B) Correlation of the two biological replicates for each genotype and time point. (A) ChIP-sequencing sample reads and mapping metrics. Inherent BMAL1 recruitment to chromatin in the liver. ![]() (A) Expression of other NAD + metabolism-related enzymes and Sirt1 from RNA-sequencing.įigure S5. Clock regulation of hepatic NAD + metabolism is autonomous and non-autonomous. Pathway names for WT are shown in gray and black for Liver-RE to highlight coherent, autonomously oscillating pathways.įigure S4. (G) Integrated pathway enrichment analysis for oscillating metabolite and oscillating transcripts in each genotype. (F) Pie chart showing the phase and amplitude of autonomously oscillating transcripts in Epidermis-RE compared to WT. (E) Enrichr was used to probe the enrichment of oscillating transcripts in each distinct group for enrichment of transcription factor target genes from published mouse liver ChIP-seq datasets (chEA). (D) Pie chart showing the phase and amplitude of autonomously oscillating transcripts in Liver-RE compared to WT. Energy expenditure group averages over 2 days (left) and quantification of light and dark periods (right) Two-way ANOVA, **=p0.01, Liver-RE vs KO – p<0.01. (D) Food intake for individual mice over a 2 day period in LD. (C) Locomotor activity traces for individual mice (double plotted for visualization), 7 days in LD followed by 7 days in DD. Right – quantification of 6 random fields of view analyzed per animal, between 15 nuclei per genotype n=3. (B) Left – BMAL1 immunostaining of liver sections co-stained with nuclear marker DRAQ7, representative maximum intensity Z projections of 15μm tissue sections are displayed. ![]() (A) Western blot for BMAL1 in various tissues from WT, KO and Liver-RE mice demonstrating the tissue-specificity of Alfp-cre in the BMAL1-Stop-FL model. Reconstitution of the liver clock using BMAL1-Stop-FL mice. ![]()
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