(A) Schematic illustrating the strategy used for generating HepKO mice. (B) Relative gene expression of Klf10 mRNA across various tissue in WT and HepKO mice as measured at ZT9 (left) and in WT and …
Values and statistical test results for Figure 1B and D and Figure 1—figure supplement 1A.
(A) Body weights (left) and daily food intake (right) of WT and HepKO mice of WT and HepKO mice (mean ± SEM, n = 44 for body weight and n = 6–8 for food intake). (B) Representative actograms of …
(A) Schematic illustrating the workflow used to assess the circadian transcriptomes of livers in WT and HepKO mice. (B) Number of rhythmic transcripts detected in the livers from WT and HepKO mice. …
MetaCycle analysis of the hepatic circadian transcriptome of WT and HepKO mice fed ad libitum and entrained in a 12:12 light/dark (LD) cycle.
(.xlsx 9.65 MB).
Phase Set Enrichment Analysis (PSEA) of the hepatic transcriptome of WT and HepKO mice fed ad libitum and entrained in a 12:12 light/dark (LD) cycle.
(A) Gene expression profiles of clock genes in the livers of WT and HepKO mice (n = 3 pools of liver per time point). (B) Gene expression profiles of genes representative of the heatmaps in Figure 2F…
Values and statistical test results for Figure 2—figure supplement 1B-F.
Expression of the Klf10 mRNA in primary WT mouse hepatocytes challenged with low (5 mM) or high (25 mM) glucose (Glu) in the absence or presence of 5 mM fructose (Fru) (mean ± SEM, n = 6) …
Values and statistical test results for Figure 3.
(A) Schematic illustrating the design of the sugar-sweetened water (SSW) challenge experiment. (B) Gene expression of Klf10 at ZT9 and ZT15 in livers of WT and HepKO mice given a chow or chow + SSW …
Values and statistical test results for Figure 3.
(A) Daily caloric intake in WT and HepKO mice under the chow and chow + sugar-sweetened water (SSW) dietary conditions (mean ± SEM, n = 8–14). (B) Blood triglyceride content at ZT15 in WT and HepKO …
(A) Expression profiles of Slc2a4 and Pklr. (B) Expression profile of Pck1. (C) Expression profiles of the Fasn and Elovl6. (A–C) Expression was determined at ZT9 and ZT15 in the liver of WT and …
Values and statistical test results for Figure 5.
(A) Expression of Slc2a2 and Slc2a5. (B) Expression of Gck, Khk, and G6pc. (C) Expression of Acaca, Acly, Me1, and Thrsp. (A–C) Expression determined at ZT15 in WT and HepKO mice given a chow or …
(A) Schematic illustrating the design of the in vitro sugar challenge experiment in WT and HepKO hepatocytes treated with low glucose (LG) or high glucose and fructose (HGF). (B) Representative …
Differentially expressed genes in WT and HepKO hepatocytes challenged with high sugar compared to unchallenged cells.
Kyoto Encyclopedia of Genes and Genomes-enriched pathway in WT and HepKO hepatocytes challenged with high sugar compared to unchallenged cells.
Values and statistical test results for Figure 6E and Figure 6—figure supplement 1.
Expression of Elovl6 in WT and HepKO primary hepatocytes treated with low glucose (LG) or high glucose and fructose (HGF) (mean ± SEM, n = 3). Statistics: nonparametric Kruskal–Wallis test. **p<0.01;…
(A) Schematic illustrating the workflow used to identify KLF10 bound loci in mouse liver at ZT9. (B) Jaspar logo representing the KLF10 response element DNA sequence and distribution of KLF10 …
Genes bound by KLF10 in their –10/+1 kb region or differentially expressed in WT vs. HepKO hepatocytes challenged with high sugar.
Cytoscape file used to generate, visualize, and analyze the KLF10 regulated metabolic network.
Values and statistical test results for Figure 7E and F.
(Top) Immunoblot showing the oscillation of KLF10 protein abundance in the liver of mice entrained in a light/dark (LD) 12:12 cycle. (Bottom) Quantification and cosinor analysis of two independent …
Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
---|---|---|---|---|
Genetic reagent (Mus musculus, C57BL/6) | WT | Weng et al., 2017 | Klf10flox/flox controls; used males for experiments | |
Genetic reagent (Mus musculus, C57BL/6) | Alb-CreERT2 | Schuler et al., 2004 | ||
Genetic reagent (Mus musculus, C57BL/6) | hepKO | This study | Used males for experiments | |
Cell line (Mus musculus) | WT | This study | Primary heaptocytes from males; used 150,000–250,000 cells per plate as indicated | |
Cell line (Mus musculus) | hepKO | This study | Primary hepatocytes from males; used 150,000–250,000 cells per plate as indicated | |
Biological sample (Mus musculus) | Liver, heart, kidney, spleen, lung, white adipose tissue | This study | ||
Biological sample (Mus musculus) | Blood | This study | ||
Antibody | Anti-KLF10 (mouse monoclonal) | CDI Laboratories | Cat. #m14-355 | WB (1:800) |
Antibody | Anti-EGF1a (mouse monoclonal) | Upstate signaling solutions | Cat. #05-235 | WB (1:1000) |
Antibody | Anti-IgG (goat polyclonal) | Sigma | Cat. #A4416 | WB (1:40,000) |
Commercial assay or kit | Glucose Hexokinase Assay Kit | Sigma | Cat. #GAHK20-1KT | |
Commercial assay or kit | PowerUp SYBR green Master Mix | Applied Biosystems | Cat. #A25779 | |
Commercial assay or kit | QIAquick PCR Purification Kit | Qiagen | Cat. #2810 | |
Commercial assay or kit | Triglycerides colorimetric assay kit | Cayman | Cat. #10010303 | |
Commercial assay or kit | NucleoSpin RNA | Macherey-Nagel | Cat. #740955250 | |
Commercial assay or kit | RNeasy mini kit | QIAGEN | Cat. #74104 | |
Commercial assay or kit | NEBNext Ultra RNA Library Prep Kit for Illumina | New England Biolabs | Cat. #E7530S | |
Commercial assay or kit | QubitTM dsDNA HS Assay Kit | Invitrogen | Cat. #Q32854 | |
Commercial assay or kit | Glucose Uptake-Glo Assay | Promega | Cat. #J1341 | |
Commercial assay or kit | Mouse Insulin Elisa | Mercodia | Cat. #10-1247-01 | |
Commercial assay or kit | Glucose (GOD-PAP) | Randox | Cat. #8318 | |
Chemical compound, drug | DMEM, low glucose, GlutaMAX Supplement, pyruvate | Gibco | Cat. #21885025 | |
Chemical compound, drug | DMEM, high glucose, GlutaMAX Supplement, pyruvate | Gibco | Cat. #10569010 | |
Chemical compound, drug | DMEM, glucose-free | Gibco | Cat. #11966025 | |
Chemical compound, drug | William’s E medium | Gibco | Cat. #22551022 | |
Chemical compound, drug | Dexamethasone | Sigma | Cat. #D0700000 | Used 10 nM final concentration |
Chemical compound, drug | Insulin-selenium-transferrin mix | Gibco | Cat. #41400045 | Used 1× final concentration |
Chemical compound, drug | Insulin (human, recombinant zinc) | Thermo Fisher Scientific | Cat. #12585014 | Used 860 nM final concentration |
Chemical compound, drug | Percoll | GE Healthcare | Cat. #17089101 | |
Chemical compound, drug | Protein G Dynabeads | Thermo Fisher Scientific | Cat. #10003D | |
Chemical compound, drug | Complete protease inhibitor cocktail | Roche | Cat. #11836153001 | |
Chemical compound, drug | Collagenase CLS3 | Worthington | Cat. #WOLS04180 | Used 500 U/mL final concentration |
Chemical compound, drug | Collagenase CLS4 | Worthington | Cat. #WOLS04186 | Used 500 U/mL final concentration |
Chemical compound, drug | Amyloglucosidase | Sigma | Cat. #11202332001 | Used 2 mg/mL final concentration |
Chemical compound, drug | SuperScript II Reverse Transcriptase | Invitrogen | Cat. #18064014 | Used 200 U per reaction |
Chemical compound, drug | Micrococcal nuclease | Thermo Fisher Scientific | Cat. #88216 | Used 80 U per reaction |
Chemical compound, drug | Protein G Dynabeads | Thermo Fisher Scientific | Cat. #10003D | |
Software, algorithm | R | Bioconductor | https://www.bioconductor.org/ | |
Software, algorithm | Trimmomatic | Bolger et al., 2014 | http://www.usadellab.org/cms/ | |
Software, algorithm | FastQC | Babraham Institute | https://www.bioinformatics.babraham.ac.uk/projects/fastqc/ | |
Software, algorithm | FASTQ Groomer | Galaxy project | https://sourceforge.net/projects/fastqgroomer/ | |
Software, algorithm | Bowtie2 | Langmead and Salzberg, 2012 | https://github.com/BenLangmead/bowtie2 | RRID: SCR_016368 |
Software, algorithm | BEDtools | Quinlan and Hall, 2010 | https://github.com/arq5x/bedtools2 | RRID: SCR_006646 |
Software, algorithm | Rsubread (featureCounts) | Liao et al., 2014 | https://git.bioconductor.org/packages/Rsubread | RRID: SCR_009803 |
Software, algorithm | MetaCycle | Wu et al., 2016 | https://github.com/gangwug/MetaCycleApp | |
Software, algorithm | Phase Set Enrichment Analysis | Zhang et al., 2016 | https://github.com/ranafi/PSEA | |
Software, algorithm | MACS2 | Feng et al., 2012 | https://pypi.org/project/MACS2/ | |
Software, algorithm | GREAT | McLean et al., 2010 | http://great.stanford.edu/public/html/splash.php | |
Software, algorithm | FIMO | Bailey et al., 2015 | http://meme-suite.org/tools/fimo | |
Software, algorithm | FastHeinz | Beisser et al., 2010 | https://www.bioconductor.org/packages/release/bioc/html/BioNet.html | |
Software, algorithm | clusterMaker2 | Morris et al., 2011 | http://www.rbvi.ucsf.edu/cytoscape/clusterMaker2/ | |
Software, algorithm | Bingo | Maere et al., 2005 | https://github.com/cytoscape/BiNGO | |
Software, algorithm | Cytoscape | Shannon et al., 2003 | https://cytoscape.org/ | |
Other | Qubit fluorometer | Thermo Fisher Scientific | Cat. #Q33238 | |
Other | FreeStyle InsuLinx glucometer | Abbott | N/A | |
Other | Laboratory rodent chow diet | Safe diets | Cat. #R03-25 | |
Other | Apistar syrup (organic) | ICKO | Cat. #HC406 |
Primers used in the study.