Journal: Cell metabolism

Article Title: FGF21 Regulates Metabolism Through Adipose-Dependent and -Independent Mechanisms

doi: 10.1016/j.cmet.2017.03.005

Figure Lengend Snippet: (A) Klb mRNA levels in epididymal WAT (eWAT), subcutaneous WAT (scWAT), interscapular BAT, and liver of 12–13 week old wild-type (WT) and β-klotho adipose-specific knockout mice (KLB AdipoKO) (n = 6/group). (B) Western blot analysis of phospho-ERK1/2 and total ERK1/2 levels from eWAT of 12–13 week old WT and KLB AdipoKO mice administered vehicle (Veh) or FGF21 (1 mg/kg) for 15 minutes. (C) Percent change in plasma glucose levels in lean, 12–14 week old, male KLB AdipoKO mice and WT littermate controls co-injected with insulin and either vehicle or FGF21 (1 mg/kg) (n = 6/group). (D) Percent change in plasma glucose levels in 16–18 week old male, diet-induced obese (DIO) KLB AdipoKO mice and WT littermate controls on co-injected with insulin and either vehicle or FGF21 (1 mg/kg) (n = 5-6/group). Plasma adiponectin levels at the indicated time points in (E) 12–13 week old, chow-fed male WT (KLBfl/fl) and (F) DIO 16–18 week old male WT (KLBfl/fl) mice on high fat diet for 12 weeks after i.p. administration of vehicle or FGF21 (1 mg/kg) (n = 8/group). (G) Percent change in plasma glucose levels in 12–14 week old, male WT (+/+ and +/−) and Adiponectin knockout (Adipoq KO) littermates co-injected with insulin and either vehicle or FGF21 (1 mg/kg) (n = 6/group). Values are mean +/− SEM. (*, P< 0.05; **, P<0.01; ***, P< 0.005; and #, P<0.001).

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies β-actin Sigma #A5316, RRID:AB_476743 phospho-ERK1/2 Cell Signaling #9101, RRID:AB_331646 total ERK1/2 Cell Signaling #9102, RRID:AB_330744 Myc Millipore #05–724, RRID:AB_309938 β-klotho R&D Systems #AF2619, AB_2131932 Chemicals, Peptides, and Recombinant Proteins Recombinant Human FGF21 Novo Nordisk N/A Human Insulin Sigma I9278 Critical Commercial Assays Human FGF21 ELISA Biovendor #RD191108200R Mouse Leptin ELISA Millipore #EZML-82K Mouse Insulin ELISA Crystal Chem #90080 Mouse Adiponectin ELISA Millipore #EZMADP-60K Glucose Autokit Wako Chemicals #439-90901 Triglycerides reagent Thermo Scientific TR22421 Total Cholesterol reagent Thermo Scientific TR13421 Experimental Models: Cell Lines Primary white adipocytes This paper N/A Experimental Models: Organisms/Strains Adiponectin KO (B6;129- Adipoq tm1Chan /J) The Jackson Laboratory Stock No: 008195 KLB fl/fl (Klb tm1.1Sakl ) Dr. Steven Kliewer, UTSW N/A Adiponectin-Cre (B6;FVB-Tg(Adipoq-cre)1Evdr/J) The Jackson Laboratory Stock No: 010803 Adiponectin-rtTA Dr. Philipp Scherer, UTSW N/A IR fl/fl (B6.129S4(FVB)- Insr tm1Khn /J) The Jackson Laboratory Stock No: 006955 TRE-caFGFR1 Dr. David Ornitz, Washu N/A UCP1-Cre mice (B6.FVB-Tg(Ucp1-cre)1Evdr/J) The Jackson Laboratory Stock No: 024670 Oligonucleotides Primers – Klb : 5′-GATGAAGAATTTCCTAAACCAGGTT-3′, 5-AACCAAACACGCGGATTTC-3′ This paper N/A Primers – Egr1 : 5′-CGAGCGAACAACCCTATGAG-3′, 5′-CATTATTCAGAGCGATGTCAGAAA-3′ This paper N/A Primers – cFos : 5′-GGACAGCCTTTCCTACTACCATTCC-3′, 5′-AAAGTTGGCACTAGAGACGGACAGA-3′ This paper N/A Primers – Bmp8b : 5′-TCAACACAACCCTCCACATCA-3′, 5′-AGATCGGAGCGTCTGAAGATC-3′ This paper N/A Primers – Zbtb16 : 5′-CCCAGTTCTCAAAGGAGGATG-3′, 5′-TTCCCACACAGCAGACAGAAG-3′ This paper N/A Primers – Fgfr1ca : 5′-TTCATCCTGGTGGTGGCAGC-3′, 5′-GCTCTTCTTGGTGCCGCTCT-3′ This paper N/A Primers – U36B4 : 5′-CGTCCTCGTTGGAGTGACA-3′, 5′-CGGTGCGTCAGGGATTG-3′ This paper N/A Software and Algorithms Graphpad Prism 7 Graphpad http://www.graphpad.com/scientific-software/prism/ PMOD version 3.709 PMOD Technologies http://www.pmod.com/web/ Inveon Research Workplace Inveon https://www.healthcare.siemens.com/molecular-imaging/preclinical-imaging/inveon-workplace/inveon-research-workplace Other N/A

Techniques: Knock-Out, Western Blot, Clinical Proteomics, Injection

Journal: Cell metabolism

Article Title: FGF21 Regulates Metabolism Through Adipose-Dependent and -Independent Mechanisms

doi: 10.1016/j.cmet.2017.03.005

Figure Lengend Snippet: (A–J) 16–18 week old DIO WT and Adipoq KO mice were administered vehicle or FGF21 (1 mg/kg) via i.p. injection daily for 3 weeks (n = 5–9/group). (A) Plasma adiponectin levels, (B) Adipoq mRNA levels in the indicated tissues, (C) daily percent change in body weight, (D) final percent change in body weight, (E) percent fat mass, (F) hepatic triglyceride levels, (G) plasma cholesterol, (H) plasma insulin and (I) plasma leptin levels in DIO WT and Adipoq KO mice administered vehicle or FGF21 for 3 weeks. (J) Insulin tolerance tests in DIO WT and Adipoq KO mice after 3 weeks (n = 5–7/group). Values are mean +/− SEM. (*, P< 0.05; **, P<0.01; ***, P< 0.005; and #, P<0.001).

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies β-actin Sigma #A5316, RRID:AB_476743 phospho-ERK1/2 Cell Signaling #9101, RRID:AB_331646 total ERK1/2 Cell Signaling #9102, RRID:AB_330744 Myc Millipore #05–724, RRID:AB_309938 β-klotho R&D Systems #AF2619, AB_2131932 Chemicals, Peptides, and Recombinant Proteins Recombinant Human FGF21 Novo Nordisk N/A Human Insulin Sigma I9278 Critical Commercial Assays Human FGF21 ELISA Biovendor #RD191108200R Mouse Leptin ELISA Millipore #EZML-82K Mouse Insulin ELISA Crystal Chem #90080 Mouse Adiponectin ELISA Millipore #EZMADP-60K Glucose Autokit Wako Chemicals #439-90901 Triglycerides reagent Thermo Scientific TR22421 Total Cholesterol reagent Thermo Scientific TR13421 Experimental Models: Cell Lines Primary white adipocytes This paper N/A Experimental Models: Organisms/Strains Adiponectin KO (B6;129- Adipoq tm1Chan /J) The Jackson Laboratory Stock No: 008195 KLB fl/fl (Klb tm1.1Sakl ) Dr. Steven Kliewer, UTSW N/A Adiponectin-Cre (B6;FVB-Tg(Adipoq-cre)1Evdr/J) The Jackson Laboratory Stock No: 010803 Adiponectin-rtTA Dr. Philipp Scherer, UTSW N/A IR fl/fl (B6.129S4(FVB)- Insr tm1Khn /J) The Jackson Laboratory Stock No: 006955 TRE-caFGFR1 Dr. David Ornitz, Washu N/A UCP1-Cre mice (B6.FVB-Tg(Ucp1-cre)1Evdr/J) The Jackson Laboratory Stock No: 024670 Oligonucleotides Primers – Klb : 5′-GATGAAGAATTTCCTAAACCAGGTT-3′, 5-AACCAAACACGCGGATTTC-3′ This paper N/A Primers – Egr1 : 5′-CGAGCGAACAACCCTATGAG-3′, 5′-CATTATTCAGAGCGATGTCAGAAA-3′ This paper N/A Primers – cFos : 5′-GGACAGCCTTTCCTACTACCATTCC-3′, 5′-AAAGTTGGCACTAGAGACGGACAGA-3′ This paper N/A Primers – Bmp8b : 5′-TCAACACAACCCTCCACATCA-3′, 5′-AGATCGGAGCGTCTGAAGATC-3′ This paper N/A Primers – Zbtb16 : 5′-CCCAGTTCTCAAAGGAGGATG-3′, 5′-TTCCCACACAGCAGACAGAAG-3′ This paper N/A Primers – Fgfr1ca : 5′-TTCATCCTGGTGGTGGCAGC-3′, 5′-GCTCTTCTTGGTGCCGCTCT-3′ This paper N/A Primers – U36B4 : 5′-CGTCCTCGTTGGAGTGACA-3′, 5′-CGGTGCGTCAGGGATTG-3′ This paper N/A Software and Algorithms Graphpad Prism 7 Graphpad http://www.graphpad.com/scientific-software/prism/ PMOD version 3.709 PMOD Technologies http://www.pmod.com/web/ Inveon Research Workplace Inveon https://www.healthcare.siemens.com/molecular-imaging/preclinical-imaging/inveon-workplace/inveon-research-workplace Other N/A

Techniques: Injection, Clinical Proteomics

Journal: Cell metabolism

Article Title: FGF21 Regulates Metabolism Through Adipose-Dependent and -Independent Mechanisms

doi: 10.1016/j.cmet.2017.03.005

Figure Lengend Snippet: (A–H) 16–18 week old DIO WT and KLB AdipoKO were implanted with osmotic minipumps delivering vehicle or FGF21 (1 mg/kg/day) for 2 weeks. (A) Plasma human FGF21, (B) plasma adiponectin, (C) final percent change in body weight, (D) hepatic triglycerides, (E) plasma triglycerides, (F) plasma glucose, (G) plasma insulin, and (H) plasma leptin levels in the indicated mice (n = 5–7/group). (I) Glucose tolerance tests in DIO WT and KLB AdipoKO mice after 2 weeks daily i.p. injection of vehicle or FGF21 (1 mg/kg) (n = 5–8/group). (J) Area under the curve (AUC) for mice in (I). (K) Insulin tolerance tests in DIO WT and KLB AdipoKO mice after 3 weeks daily i.p. injection of vehicle or FGF21 (1 mg/kg) (n = 5–8/group). (L) Area under the curve for mice in (K). (M) caFGFR1 mRNA levels in eWAT, scWAT, and BAT of WT, Adipo-rtTA transgenic, TRE-caFGFR1 transgenic, and Adipo-rtTA/TRE-caFGFR1 double transgenic mice provided water with or without doxycycline (DOX) for 5 days. (N–S) WT, TRE-caFGFR1 transgenic, and Adipo-rtTA/TRE-caFGFR1 double transgenic mice were induced to obesity by 12 weeks of HFD feeding. Groups of mice were then switched to HFD + DOX for 2 weeks (n = 7–10/group). (N) Western blot analysis of Myc and phospho-ERK1/2 levels in eWAT from mice with the indicated genotypes. (O) Egr1 and cFos mRNA expression in eWAT of the indicated mice. (P) Final percent body weight change, (Q) hepatic triglycerides, (R) plasma insulin, and (S) plasma glucose levels in the indicated mice on HFD + DOX for 2 weeks. Values are mean +/− SEM. (*, P< 0.05; **, P<0.01; ***, P< 0.005; and #, P<0.001).

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies β-actin Sigma #A5316, RRID:AB_476743 phospho-ERK1/2 Cell Signaling #9101, RRID:AB_331646 total ERK1/2 Cell Signaling #9102, RRID:AB_330744 Myc Millipore #05–724, RRID:AB_309938 β-klotho R&D Systems #AF2619, AB_2131932 Chemicals, Peptides, and Recombinant Proteins Recombinant Human FGF21 Novo Nordisk N/A Human Insulin Sigma I9278 Critical Commercial Assays Human FGF21 ELISA Biovendor #RD191108200R Mouse Leptin ELISA Millipore #EZML-82K Mouse Insulin ELISA Crystal Chem #90080 Mouse Adiponectin ELISA Millipore #EZMADP-60K Glucose Autokit Wako Chemicals #439-90901 Triglycerides reagent Thermo Scientific TR22421 Total Cholesterol reagent Thermo Scientific TR13421 Experimental Models: Cell Lines Primary white adipocytes This paper N/A Experimental Models: Organisms/Strains Adiponectin KO (B6;129- Adipoq tm1Chan /J) The Jackson Laboratory Stock No: 008195 KLB fl/fl (Klb tm1.1Sakl ) Dr. Steven Kliewer, UTSW N/A Adiponectin-Cre (B6;FVB-Tg(Adipoq-cre)1Evdr/J) The Jackson Laboratory Stock No: 010803 Adiponectin-rtTA Dr. Philipp Scherer, UTSW N/A IR fl/fl (B6.129S4(FVB)- Insr tm1Khn /J) The Jackson Laboratory Stock No: 006955 TRE-caFGFR1 Dr. David Ornitz, Washu N/A UCP1-Cre mice (B6.FVB-Tg(Ucp1-cre)1Evdr/J) The Jackson Laboratory Stock No: 024670 Oligonucleotides Primers – Klb : 5′-GATGAAGAATTTCCTAAACCAGGTT-3′, 5-AACCAAACACGCGGATTTC-3′ This paper N/A Primers – Egr1 : 5′-CGAGCGAACAACCCTATGAG-3′, 5′-CATTATTCAGAGCGATGTCAGAAA-3′ This paper N/A Primers – cFos : 5′-GGACAGCCTTTCCTACTACCATTCC-3′, 5′-AAAGTTGGCACTAGAGACGGACAGA-3′ This paper N/A Primers – Bmp8b : 5′-TCAACACAACCCTCCACATCA-3′, 5′-AGATCGGAGCGTCTGAAGATC-3′ This paper N/A Primers – Zbtb16 : 5′-CCCAGTTCTCAAAGGAGGATG-3′, 5′-TTCCCACACAGCAGACAGAAG-3′ This paper N/A Primers – Fgfr1ca : 5′-TTCATCCTGGTGGTGGCAGC-3′, 5′-GCTCTTCTTGGTGCCGCTCT-3′ This paper N/A Primers – U36B4 : 5′-CGTCCTCGTTGGAGTGACA-3′, 5′-CGGTGCGTCAGGGATTG-3′ This paper N/A Software and Algorithms Graphpad Prism 7 Graphpad http://www.graphpad.com/scientific-software/prism/ PMOD version 3.709 PMOD Technologies http://www.pmod.com/web/ Inveon Research Workplace Inveon https://www.healthcare.siemens.com/molecular-imaging/preclinical-imaging/inveon-workplace/inveon-research-workplace Other N/A

Techniques: Clinical Proteomics, Injection, Transgenic Assay, Western Blot, Expressing

Journal: Cell metabolism

Article Title: FGF21 Regulates Metabolism Through Adipose-Dependent and -Independent Mechanisms

doi: 10.1016/j.cmet.2017.03.005

Figure Lengend Snippet: (A) Klb mRNA levels in eWAT, scWAT, BAT, and liver of 12–14 week old WT and β-klotho brown adipose-specific knockout mice (KLB BatKO) (n = 6/group). (B) Percent change in plasma glucose levels in 12–14 week old, male KLB BatKO mice and WT controls co-injected with insulin and either vehicle or FGF21 (1 mg/kg) (n = 6/group). (C) Percent change in plasma glucose levels in 12–14 week old, male adipose-specific insulin receptor knockout (IR AdipoKO) mice and WT controls co-injected with insulin and either vehicle or FGF21 (1 mg/kg) (n = 5–7/group). (D) PET/CT images of relative [18F]-FDG uptake in WT and KLB BatKO mice co-injected with insulin and either vehicle or FGF21 (1 mg/kg). (E) Average standard uptake value in interscapular BAT of the indicated mice (n = 5–8/group). (F) Tissue-specific glucose uptake assays using [3H]-2-deoxyglucose in WT and KLB BatKO mice co-injected with insulin and either vehicle or FGF21 (1 mg/kg) for 90 minutes (n = 5–13/group). Values are mean +/− SEM. (*, P< 0.05; **, P<0.01; ***, P< 0.005; and #, P<0.001).

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies β-actin Sigma #A5316, RRID:AB_476743 phospho-ERK1/2 Cell Signaling #9101, RRID:AB_331646 total ERK1/2 Cell Signaling #9102, RRID:AB_330744 Myc Millipore #05–724, RRID:AB_309938 β-klotho R&D Systems #AF2619, AB_2131932 Chemicals, Peptides, and Recombinant Proteins Recombinant Human FGF21 Novo Nordisk N/A Human Insulin Sigma I9278 Critical Commercial Assays Human FGF21 ELISA Biovendor #RD191108200R Mouse Leptin ELISA Millipore #EZML-82K Mouse Insulin ELISA Crystal Chem #90080 Mouse Adiponectin ELISA Millipore #EZMADP-60K Glucose Autokit Wako Chemicals #439-90901 Triglycerides reagent Thermo Scientific TR22421 Total Cholesterol reagent Thermo Scientific TR13421 Experimental Models: Cell Lines Primary white adipocytes This paper N/A Experimental Models: Organisms/Strains Adiponectin KO (B6;129- Adipoq tm1Chan /J) The Jackson Laboratory Stock No: 008195 KLB fl/fl (Klb tm1.1Sakl ) Dr. Steven Kliewer, UTSW N/A Adiponectin-Cre (B6;FVB-Tg(Adipoq-cre)1Evdr/J) The Jackson Laboratory Stock No: 010803 Adiponectin-rtTA Dr. Philipp Scherer, UTSW N/A IR fl/fl (B6.129S4(FVB)- Insr tm1Khn /J) The Jackson Laboratory Stock No: 006955 TRE-caFGFR1 Dr. David Ornitz, Washu N/A UCP1-Cre mice (B6.FVB-Tg(Ucp1-cre)1Evdr/J) The Jackson Laboratory Stock No: 024670 Oligonucleotides Primers – Klb : 5′-GATGAAGAATTTCCTAAACCAGGTT-3′, 5-AACCAAACACGCGGATTTC-3′ This paper N/A Primers – Egr1 : 5′-CGAGCGAACAACCCTATGAG-3′, 5′-CATTATTCAGAGCGATGTCAGAAA-3′ This paper N/A Primers – cFos : 5′-GGACAGCCTTTCCTACTACCATTCC-3′, 5′-AAAGTTGGCACTAGAGACGGACAGA-3′ This paper N/A Primers – Bmp8b : 5′-TCAACACAACCCTCCACATCA-3′, 5′-AGATCGGAGCGTCTGAAGATC-3′ This paper N/A Primers – Zbtb16 : 5′-CCCAGTTCTCAAAGGAGGATG-3′, 5′-TTCCCACACAGCAGACAGAAG-3′ This paper N/A Primers – Fgfr1ca : 5′-TTCATCCTGGTGGTGGCAGC-3′, 5′-GCTCTTCTTGGTGCCGCTCT-3′ This paper N/A Primers – U36B4 : 5′-CGTCCTCGTTGGAGTGACA-3′, 5′-CGGTGCGTCAGGGATTG-3′ This paper N/A Software and Algorithms Graphpad Prism 7 Graphpad http://www.graphpad.com/scientific-software/prism/ PMOD version 3.709 PMOD Technologies http://www.pmod.com/web/ Inveon Research Workplace Inveon https://www.healthcare.siemens.com/molecular-imaging/preclinical-imaging/inveon-workplace/inveon-research-workplace Other N/A

Techniques: Knock-Out, Clinical Proteomics, Injection, Positron Emission Tomography-Computed Tomography