nampt inhibitor fk866  (Selleck Chemicals)

Journal: Toxins

Article Title: Nicotinamide Ameliorates Deoxynivalenol-Induced Injury in Renal Cells via Inhibiting PARP1 Hyperactivation and Restoring NAD + Homeostasis

doi: 10.3390/toxins18050227

Figure Lengend Snippet: ( A ) Serum NAD + content in Duroc × Landrace × Yorkshire three-way crossbred piglets after 28 days of DON challenge (2 mg/kg diet), n = 6 per group. ( B ) Intracellular NAD + levels in HEK293T cells treated with 200 ng/mL DON for 24 h. ( C ) Protective effect of exogenous NAD + (0, 0.5, 1, 50, 500 μg/mL) on DON-induced reduction in HEK293T cell viability (200 ng/mL DON for 24 h), detected by CCK-8 assay. ( D ) Purified recombinant human NAMPT was incubated with 100 μM DON or equal-volume DMSO (vehicle control) at 4 °C for 1.5 h. Aliquots were heated at the indicated temperature gradient for 3 min, and centrifuged to remove precipitated denatured proteins. Soluble NAMPT in supernatants was analyzed by Western blot. The line graph shows the normalized grayscale analysis with unheated protein serving as the internal standard. ( E ) Molecular docking model of DON and human NAMPT (PDB ID: 4KFN) indicates the key binding sites (Arg-196 and Asp-219) in the NAMPT active site. Statistical significance was designated as * p < 0.05, ** p < 0.01, and *** p < 0.001; ns = no significant difference.

Article Snippet: Primers targeting the full-length CDS of human NAMPT (GenBank accession No. NM_005746.3 ) were synthesized by Sangon Biotech ( ).

Techniques: CCK-8 Assay, Purification, Recombinant, Incubation, Control, Western Blot, Binding Assay

Journal: Inflammation

Article Title: Adipose-Derived Proinflammatory Visfatin Promotes Vascular Calcification via TLR4 and is Suppressed by Empagliflozin

doi: 10.1007/s10753-026-02507-5

Figure Lengend Snippet: Empagliflozin suppresses the production and release of visfatin in adipose tissue and adipocytes. ( A ) Serum visfatin levels measured by ELISA ( n = 6). ( B ) Western blot analysis of visfatin levels in the serum of mice with vascular calcification and vascular calcification + empagliflozin gavage ( n = 6). ( C ) RT‒qPCR was used to assess the mRNA levels of visfatin in visceral adipose tissue ( n = 6). ( D ) Immunohistochemical staining of visfatin in the epididymal adipose tissue of mice with vascular calcification and vascular calcification + empagliflozin gavage ( n = 6). Scale bar: 50 μm. ( E ) Visfatin levels in the culture supernatant were measured by ELISA ( n = 6). F‒G. The protein and mRNA levels of visfatin in differentiated 3T3-L1 adipocytes ( n = 6). * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: Empagliflozin (HY-15409), BAY 11–7085 (HY-10257), insulin (HY-P0035), rosiglitazone (HY-17386), and visfatin (HY-P701314) were purchased from MedChemExpress (New Jersey, USA), and FK866 (658084-64-1) was purchased from Selleck Chemicals (Shanghai, China).

Techniques: Enzyme-linked Immunosorbent Assay, Western Blot, Immunohistochemical staining, Staining

Journal: Inflammation

Article Title: Adipose-Derived Proinflammatory Visfatin Promotes Vascular Calcification via TLR4 and is Suppressed by Empagliflozin

doi: 10.1007/s10753-026-02507-5

Figure Lengend Snippet: Effect of the p38 MAPK pathway on the empagliflozin-mediated regulation of visfatin expression and secretion in adipocytes. ( A ) Western blot analysis was used to evaluate the effect of calcification medium on the activation of JNK, ERK, and p38 MAPK in adipocytes. β-actin was used as a loading control ( n = 5). ( B ) Adipocytes were treated with SP600125 (a JNK inhibitor), PD098059 (an ERK1/2 inhibitor), or SB203580 (a p38 inhibitor) during exposure to calcification medium. Western blot analysis was used to measure the protein levels of visfatin ( n = 5). ( C ) RT–qPCR was used to assess the mRNA level of visfatin ( n = 6). ( D ) After they were exposed to calcification medium, adipocytes were treated with SB203580 (a p38 inhibitor), and Western blotting was used to measure the protein levels of visfatin in adipocytes ( n = 6). ( E ) After treatment with calcification medium, the effect of empagliflozin on the phosphorylation of p38 in adipocytes was assessed by Western blot analysis. β-actin was used as a loading control ( n = 5). * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: Empagliflozin (HY-15409), BAY 11–7085 (HY-10257), insulin (HY-P0035), rosiglitazone (HY-17386), and visfatin (HY-P701314) were purchased from MedChemExpress (New Jersey, USA), and FK866 (658084-64-1) was purchased from Selleck Chemicals (Shanghai, China).

Techniques: Expressing, Western Blot, Activation Assay, Control, Quantitative RT-PCR, Phospho-proteomics

Journal: Inflammation

Article Title: Adipose-Derived Proinflammatory Visfatin Promotes Vascular Calcification via TLR4 and is Suppressed by Empagliflozin

doi: 10.1007/s10753-026-02507-5

Figure Lengend Snippet: Empagliflozin inhibits the endogenous binding of NF-κB to the visfatin promoter. ( A ) Confocal microscopy of immunofluorescence staining for NF-κB in VSMCs, both in the presence and absence of calcification medium. Scale bar: 20 μm. ( B ) During exposure to calcification medium, adipocytes were treated with BAY 11–7085 (an NF-κB inhibitor), and Western blotting was used to measure the protein levels of visfatin in adipocytes ( n = 6). RT–qPCR was used to assess the mRNA level of visfatin ( n = 6). ( C ) Confocal microscopy of immunofluorescence staining for NF-κB in VSMCs following treatment of adipocytes with empagliflozin and SB203580 (p38 inhibitor). Scale bar: 20 μm. ( D ) NF-κB was overexpressed in HEK293T cells cotransfected with a plasmid carrying the visfatin promoter that drives firefly luciferase and Renilla luciferase. Firefly luciferase and Renilla luciferase activity assays were subsequently performed ( n = 3). ( E ) The sequence labeled 1 corresponds to the first exon of the mRNA, which is indicated in bold and enclosed in a box. Additionally, the sequences within the other two rectangles represent the putative NF-κB binding sites. ( F ) RT–qPCR was used to quantitatively analyze the binding of NF-κB to the visfatin promoter region ( n = 3). ( G ) After ChIP was performed, agarose gel electrophoresis revealed that NF-κB could bind to the visfatin promoter region ( n = 3). * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: Empagliflozin (HY-15409), BAY 11–7085 (HY-10257), insulin (HY-P0035), rosiglitazone (HY-17386), and visfatin (HY-P701314) were purchased from MedChemExpress (New Jersey, USA), and FK866 (658084-64-1) was purchased from Selleck Chemicals (Shanghai, China).

Techniques: Binding Assay, Confocal Microscopy, Immunofluorescence, Staining, Western Blot, Quantitative RT-PCR, Plasmid Preparation, Luciferase, Activity Assay, Sequencing, Labeling, Agarose Gel Electrophoresis