rabbit anti nampt  (Proteintech)

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: Stem Cells Translational Medicine

Article Title: METTL1-deficient mesenchymal stem cells protect against metabolic-associated fatty liver disease by increasing NAMPT secretion

doi: 10.1093/stcltm/szag016

Figure Lengend Snippet: FK866 primed METTL1-deficient MSCs fail to protect against MASLD-related metabolic disorders due to impaired NAMPT secretion. (A) Schematic diagram of the animal experiment. (B) Assessment of liver weight and the liver-to-body weight ratio in the indicated mice. (C) Evaluation of fasting blood glucose levels in the specified mice. (D) GTT and ITT analyses were conducted on the designated groups. (E) Measurement of serum ALT and AST levels was conducted 7 weeks post-cell transplantation. (F) Representative images of HE and Oil Red O staining of mouse liver tissues (Scale bar = 100 μm). (G) Determination of TG and TC levels in the liver tissues. (H) Western blot analysis was performed to investigate the proteins associated with lipid metabolism and NAMPT/SIRT1 signaling in mouse liver tissues. For all statistical graphs, individual data points represent individual mice, and data are presented as mean ±S.E.M. Statistical significance is indicated as shown in the figure.

Article Snippet: Additionally, after treating the cells with fresh medium containing the NAMPT inhibitor FK866 (S2799, Selleck, China) for 24 hours, the cells were harvested for subsequent experiments.

Techniques: Transplantation Assay, Staining, Western Blot

Journal: Stem Cells Translational Medicine

Article Title: METTL1-deficient mesenchymal stem cells protect against metabolic-associated fatty liver disease by increasing NAMPT secretion

doi: 10.1093/stcltm/szag016

Figure Lengend Snippet: METTL1-deficient MSCs exhibit elevated NAMPT secretion. (A) Volcano plot depicting differentially expressed genes between MSC shGFP and MSC shMETTL1 , as analyzed by RNA sequencing. (B) Identification of genes significantly enriched in GO analysis for MSC shMETTL1 cells. (C) Venn diagram illustrating the distinct proteins present in the secretome of MSC shGFP and MSC shMETTL1 . (D) GO analysis of proteins that are significantly enriched in the secretome of MSC shMETTL1 cells. (E) Heatmap representation of differentially expressed secretory proteins implicated in the regulation of lipid metabolism associated with MASLD in MSC shGFP and MSC shMETTL1 . (F) qPCR analysis of the expression of relevant genes in the specified cells. (G) ELISA measurements of the levels of differentially expressed secretory proteins in the supernatants of MSC shGFP and MSC shMETTL1 cells. For all statistical graphs, individual data points represent independent experimental replicates, and data are presented as mean ± S.E.M. Statistical significance is indicated as shown in the figure.

Article Snippet: Additionally, a primary antibody to METTL1 (14994-1-AP), SIRT1 (13161-1-AP), NAMPT (11776-1-AP) and SREBP1 (14088-1-AP) were purchased from Proteintech.

Techniques: RNA Sequencing, Expressing, Enzyme-linked Immunosorbent Assay

Journal: Stem Cells Translational Medicine

Article Title: METTL1-deficient mesenchymal stem cells protect against metabolic-associated fatty liver disease by increasing NAMPT secretion

doi: 10.1093/stcltm/szag016

Figure Lengend Snippet: NAMPT/SIRT1/SREBP1 mediates the protective effects of METTL1-deficient MSCs in MASLD. (A-C) Representative images of IF staining for NAMPT, SIRT1 and SREBP1 in HepG2 cells co-cultured with MSC shGFP and MSC shMETTL1 cells after treatment with FFA (Scale bar = 20 μm). (D, E) Western blot analysis of NAMPT, SIRT1 and SREBP1 expression in HepG2 cells co-cultured with MSC shGFP and MSC shMETTL1 cells following FFA treatment. (F) The NAD+ content was measured in the indicated cells. (G-I) Representative images of IF staining for NAMPT, SIRT1 and SREBP1 in liver tissues from mice transplanted with MSC shGFP and MSC shMETTL1 cells after 16 weeks of HFD feeding (Scale bar = 100 μm). (J, K) Western blot analysis of NAMPT, SIRT1, and SREBP1 expression in liver tissues from mice transplanted with MSC shGFP and MSC shMETTL1 cells after 16 weeks of HFD feeding. (L) The NAD+ content was measured in the indicated mouse liver tissues. For all statistical graphs, data are presented as mean ± S.E.M., with statistical significance indicated in the figure.

Article Snippet: Additionally, a primary antibody to METTL1 (14994-1-AP), SIRT1 (13161-1-AP), NAMPT (11776-1-AP) and SREBP1 (14088-1-AP) were purchased from Proteintech.

Techniques: Staining, Cell Culture, Western Blot, Expressing

Journal: Stem Cells Translational Medicine

Article Title: METTL1-deficient mesenchymal stem cells protect against metabolic-associated fatty liver disease by increasing NAMPT secretion

doi: 10.1093/stcltm/szag016

Figure Lengend Snippet: FK866 primed METTL1-deficient MSCs fail to protect against MASLD-related metabolic disorders due to impaired NAMPT secretion. (A) Schematic diagram of the animal experiment. (B) Assessment of liver weight and the liver-to-body weight ratio in the indicated mice. (C) Evaluation of fasting blood glucose levels in the specified mice. (D) GTT and ITT analyses were conducted on the designated groups. (E) Measurement of serum ALT and AST levels was conducted 7 weeks post-cell transplantation. (F) Representative images of HE and Oil Red O staining of mouse liver tissues (Scale bar = 100 μm). (G) Determination of TG and TC levels in the liver tissues. (H) Western blot analysis was performed to investigate the proteins associated with lipid metabolism and NAMPT/SIRT1 signaling in mouse liver tissues. For all statistical graphs, individual data points represent individual mice, and data are presented as mean ±S.E.M. Statistical significance is indicated as shown in the figure.

Article Snippet: Additionally, a primary antibody to METTL1 (14994-1-AP), SIRT1 (13161-1-AP), NAMPT (11776-1-AP) and SREBP1 (14088-1-AP) were purchased from Proteintech.

Techniques: Transplantation Assay, Staining, Western Blot

Journal: Stem Cells Translational Medicine

Article Title: METTL1-deficient mesenchymal stem cells protect against metabolic-associated fatty liver disease by increasing NAMPT secretion

doi: 10.1093/stcltm/szag016

Figure Lengend Snippet: METTL1-overexpressing MSCs accelerate MASLD progression with reduced NAMPT secretion. (A) Schematic diagram of the animal experiment. (B) GTT and ITT analyses were conducted on the designated groups. (C) Representative images of HE and Oil Red O staining of mouse liver tissues (Scale bar = 100 μm). (D) Measurement of TG and TC levels in the liver tissues. (E) qPCR analysis of lipid synthesis-related genes in the designated groups. (F) Western blot analysis was performed to investigate the proteins associated with lipid metabolism and NAMPT/SIRT1 signaling in mouse liver tissues. For all statistical analyses, individual data points represent individual mice, and data are presented as mean ±S.E.M. Statistical significance is indicated as shown in the figure.

Article Snippet: Additionally, a primary antibody to METTL1 (14994-1-AP), SIRT1 (13161-1-AP), NAMPT (11776-1-AP) and SREBP1 (14088-1-AP) were purchased from Proteintech.

Techniques: Staining, Western Blot