Journal: Molecular Metabolism

Article Title: FGF21 resistance is not mediated by downregulation of beta-klotho expression in white adipose tissue

doi: 10.1016/j.molmet.2017.03.009

Figure Lengend Snippet: Decreased expression of β-klotho in white adipose tissue correlates with elevated levels of circulating FGF21 . (A) Plasma FGF21 protein levels in male WT C57Bl/6 mice ad libitum fed chow or 60% high fat diet (HFD) for 16 weeks (n = 6–7/group). (B–G) Quantitative real-time PCR (QPCR) analysis of Klb and Fgfr1c mRNA expression in epididymal white adipose tissue (eWAT; B, C), liver (D–E), and hypothalamus (F–G) (n = 6–7/group). (H) Western blot analysis of KLB protein expression in eWAT of male WT C57Bl/6 mice ad libitum fed chow or 60% high fat diet (HFD) for 16 weeks. (I) Representative western blots of KLB protein expression in eWAT, brown adipose tissue (BAT), and liver of WT C57Bl/6 mice ad libitum fed 60% high fat diet (HFD) for the indicated time (5–6 mice/group). (J) Plasma FGF21 and insulin levels from the mice in (I). (K) Liver Fgf21 mRNA levels from mice in (I). Values are mean +/− SEM. (*, P < 0.05; **, P < 0.01; ***, P < 0.005; and #, P < 0.001 compared to WT; for (J) significance is relative to time 0).

Article Snippet: Antibody information: β-klotho (R&D Systems, #AF2619), β-actin (Sigma, #A5316), IR-β (Santa Cruz, #sc-711), phospho-FRS2α (Cell Signaling, #3864), phospho-ERK1/2 (Cell Signaling, #9101), total ERK1/2 (Cell Signaling, #9102), phospho-MEK (Cell Signaling, #9121), and total MEK (Cell Signaling, #9122).

Techniques: Expressing, Clinical Proteomics, Real-time Polymerase Chain Reaction, Western Blot

Journal: Molecular Metabolism

Article Title: FGF21 resistance is not mediated by downregulation of beta-klotho expression in white adipose tissue

doi: 10.1016/j.molmet.2017.03.009

Figure Lengend Snippet: Generation of adipose-specific β-klotho transgenic mice . (A) Schematic of inducible β-klotho transgenic construct (inKLB). (B) Western blot analysis of KLB protein expression in epididymal white adipose tissue (eWAT) of 16 week chow-fed and high fat diet (HFD)-fed wild-type (WT) mice compared to HFD fed WT and adipose-specific β-klotho transgenic (KLB AdipoTG) mice. QPCR analysis of Klb and Tdtomato mRNA expression in eWAT (C, D), inguinal white adipose tissue (iWAT; E, F), brown adipose tissue (BAT; G, H), and liver (I, J) of WT and KLB AdipoTG mice fed chow or 60% HFD for 12 weeks (n = 7–11/group). (K) Western blot analysis of cytoplasmic and membrane KLB protein expression in epididymal white adipose tissue of wild-type (WT) and KLB adipose-specific transgenic mice (KLB AdipoTG). Values are mean +/− SEM. (*, P < 0.05; ***, P < 0.005; and #, P < 0.001 compared to WT; nd = not detected).

Article Snippet: Antibody information: β-klotho (R&D Systems, #AF2619), β-actin (Sigma, #A5316), IR-β (Santa Cruz, #sc-711), phospho-FRS2α (Cell Signaling, #3864), phospho-ERK1/2 (Cell Signaling, #9101), total ERK1/2 (Cell Signaling, #9102), phospho-MEK (Cell Signaling, #9121), and total MEK (Cell Signaling, #9122).

Techniques: Transgenic Assay, Construct, Western Blot, Expressing, Membrane

Journal: Molecular Metabolism

Article Title: FGF21 resistance is not mediated by downregulation of beta-klotho expression in white adipose tissue

doi: 10.1016/j.molmet.2017.03.009

Figure Lengend Snippet: Maintenance of β-klotho expression in white adipose tissue does not improve metabolic profiles . (A) Body weight curves and (B) body composition of male WT and KLB AdipoTG mice on HFD for 12 weeks (n = 8–10/group). (C) Glucose tolerance (GTTs) and (D) insulin tolerance tests (ITTs) of lean, male WT and KLB AdipoTG mice (n = 6/group). (E) GTTs and (F) ITTs of diet-induced obese (DIO), male WT and KLB AdipoTG mice on HFD for 12 and 14 weeks, respectively (n = 6/group). (G) Hepatic triglyceride levels in DIO WT and KLB AdipoTG mice. Values are mean +/− SEM. (*, P < 0.05; ***, P < 0.005; and #, P < 0.001 compared to WT).

Article Snippet: Antibody information: β-klotho (R&D Systems, #AF2619), β-actin (Sigma, #A5316), IR-β (Santa Cruz, #sc-711), phospho-FRS2α (Cell Signaling, #3864), phospho-ERK1/2 (Cell Signaling, #9101), total ERK1/2 (Cell Signaling, #9102), phospho-MEK (Cell Signaling, #9121), and total MEK (Cell Signaling, #9122).

Techniques: Expressing

Journal: Molecular Metabolism

Article Title: FGF21 resistance is not mediated by downregulation of beta-klotho expression in white adipose tissue

doi: 10.1016/j.molmet.2017.03.009

Figure Lengend Snippet: White adipose tissue and systemic FGF21 sensitivity is not improved by preservation of β-klotho expression . (A) Plasma FGF21 and (B) hepatic Fgf21 mRNA levels in WT and KLB AdipoTG mice fed chow or HFD for 14 weeks (n = 8–9/group). (C) Western blot analysis of phospho-ERK1/2, total ERK1/2, phospho-FRS2α, phospho-MEK, and total MEK from eWAT of chow and HFD fed WT and KLB AdipoTG mice administered either vehicle or FGF21 (0.1 mg/kg) for 15 min. Values are mean +/− SEM. (#, P < 0.001 compared to WT).

Article Snippet: Antibody information: β-klotho (R&D Systems, #AF2619), β-actin (Sigma, #A5316), IR-β (Santa Cruz, #sc-711), phospho-FRS2α (Cell Signaling, #3864), phospho-ERK1/2 (Cell Signaling, #9101), total ERK1/2 (Cell Signaling, #9102), phospho-MEK (Cell Signaling, #9121), and total MEK (Cell Signaling, #9122).

Techniques: Preserving, Expressing, Clinical Proteomics, Western Blot

Journal: Gerontology

Article Title: Role of NADPH Oxidases in Renal Aging.

doi: 10.1159/000529392

Figure Lengend Snippet: Fig. 5. Representative immunoblots from renal cortical tissues in db/m, db/db mice. Representative immunoblots for TGFβ1, type IV collagen, Klotho, NOX1, NOX2, and NOX4.

Article Snippet: Proteins were transferred onto polyvinylidene difluoride membranes, which were hybridized in blocking buffer overnight at 4°C with rabbit polyclonal anti-Nox1 antibody (1:1,000, Abcam Plc, Cambridge, MA, USA), rabbit polyclonal anti-Nox2 antibody (1:1,000, Bioworld Technology, St. Louis Park, MN, USA), rabbit polyclonal anti-Nox4 antibody (1:1,000, Bioworld Technology, St. Louis Park, MN, USA), rabbit polyclonal anti-type IV collagen antibody (1:1,000, Abcam Plc, Cambridge, MA, USA), rabbit polyclonal anti-TGFβ1 antibody (1:1,000, Abbkine, Waltham, MA, USA), goat polyclonal anti-Klotho antibody (1:1,000, R&D Systems Inc., Minneapolis, MN, USA), or mouse monoclonal anti-β-actin antibody (1:5,000, Sigma-Aldrich, St. Louis, MO, USA).

Techniques: Western Blot

Journal: Gerontology

Article Title: Role of NADPH Oxidases in Renal Aging.

doi: 10.1159/000529392

Figure Lengend Snippet: Fig. 8. a Glomerulosclerosis index and scoring of F4/80 positive cells. b IHC scoring for TGFβ1, type IV collagen, and PAI-1. c Representative immunoblots for TGFβ1, type IV collagen, Klotho, NOX1, NOX2, and NOX4.

Article Snippet: Proteins were transferred onto polyvinylidene difluoride membranes, which were hybridized in blocking buffer overnight at 4°C with rabbit polyclonal anti-Nox1 antibody (1:1,000, Abcam Plc, Cambridge, MA, USA), rabbit polyclonal anti-Nox2 antibody (1:1,000, Bioworld Technology, St. Louis Park, MN, USA), rabbit polyclonal anti-Nox4 antibody (1:1,000, Bioworld Technology, St. Louis Park, MN, USA), rabbit polyclonal anti-type IV collagen antibody (1:1,000, Abcam Plc, Cambridge, MA, USA), rabbit polyclonal anti-TGFβ1 antibody (1:1,000, Abbkine, Waltham, MA, USA), goat polyclonal anti-Klotho antibody (1:1,000, R&D Systems Inc., Minneapolis, MN, USA), or mouse monoclonal anti-β-actin antibody (1:5,000, Sigma-Aldrich, St. Louis, MO, USA).

Techniques: Western Blot

Journal: FEBS Open Bio

Article Title: Interleukin‐36α as a potential biomarker for renal tubular damage induced by dietary phosphate load

doi: 10.1002/2211-5463.12845

Figure Lengend Snippet: Localization of IL‐36α expression induced by phosphate load. Wild‐type mice at 12 weeks of age were placed on a high‐phosphate diet containing 2.0% Pi for 10 days (A), 4 weeks (B), and 8 weeks (C). Serial paraffin sections of the kidneys were subjected to immunohistochemical staining using antibodies against IL‐36α, Klotho, Osteopontin, and F4/80. (A) Klotho‐positive tubules expressed IL‐36α. Osteopontin‐positive tubules expressed IL‐36α. (B, C) IL‐36α‐positive tubules expressed Osteopontin and were surrounded by F4/80‐positive cells. Arrows indicate identical renal tubule in serial sections. Bar = 50 μm.

Article Snippet: After blocking, the sections were incubated with a goat polyclonal antibody against mouse IL‐36α (diluted 1 : 100 with PBS containing 1% BSA; AF2297; R&D Systems, Minneapolis, MN, USA), a goat polyclonal antibody against mouse Klotho (diluted 1 : 100 with PBS containing 1% BSA; BAF1819; R&D Systems), a rat monoclonal antibody against mouse F4/80 (diluted 1 : 100 with PBS containing 1% BSA; GTX26640; GeneTex, San Antonio, TX, USA), or a mouse monoclonal antibody against mouse Osteopontin (diluted 1 : 100 with PBS containing 1% BSA; sc‐21742; Santa Cruz, Dallas, TX, USA) at 4 °C overnight and then with Histofine Simple Stain Mouse MAX PO (Nichirei Corporation) at room temperature for 1 h. Immunoreactivity was visualized by incubation with the DAB substrate kit (Dako; Agilent Pathology Solutions).

Techniques: Expressing, Immunohistochemical staining, Staining