sirt6 assay kit  (BPS Bioscience)

Journal: Journal of cachexia, sarcopenia and muscle

Article Title: SIRT6 Ameliorates Cancer Cachexia-Associated Adipose Wasting by Suppressing TNFR2 Signalling in Mice.

doi: 10.1002/jcsm.13734

Figure Lengend Snippet: FIGURE 1 | Serum SIRT6 concentrations are associated with cachexia in patients with cancer. (A) Diagram elucidating the collection and analy- sis of patient serum sample. Serum collected from healthy controls, cachectic cancer patients and non-cachectic cancer patients, were used to mea- sure SIRT6 concentrations by enzyme-linked immunosorbent assay (ELISA). (B) Serum SIRT6 concentrations of healthy controls (n = 22) and cancer patients (n = 22) were compared. (C) Serum SIRT6 concentrations of cachectic cancer patients (n = 12) and non-cachectic cancer patients (n = 10) were compared. **p < 0.01.

Article Snippet: Human serum SIRT6, TNFα and TNFR2 concentrations were analysed by Human NAD- dependent deacetylase sirtuin- 6 (SIRT6) ELISA kit (CSB- E17018h, CUSABIO), human tumour necrosis factor α (TNF- α) ELISA kit (CSB- E04740h, CUSABIO) and human soluble tumour necrosis factor receptor 2 (sTNF- R2) ELISA kit (CSB- E11266h, CUSABIO) according to the manufacturer's instructions, individually.

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Journal of cachexia, sarcopenia and muscle

Article Title: SIRT6 Ameliorates Cancer Cachexia-Associated Adipose Wasting by Suppressing TNFR2 Signalling in Mice.

doi: 10.1002/jcsm.13734

Figure Lengend Snippet: FIGURE 2 | SIRT6 overexpression prevents body weight loss and adipose tissue wasting in tumour-bearing mice. (A–D) SIRT6 transgenic (TG) and wild type (WT) mice were inoculated with LLC cells or PBS and euthanized 21 days after tumour injection. Tumour weight (A), carcass weight (B), weight of total fat tissue (C), weight of epididymal white adipose tissue (eWAT), inguinal white adipose tissue (iWAT) and brown adipose tissue (BAT) (D) were measured (n = 6 per group). (E) Representative H&E-stained images of eWAT, iWAT and BAT. Adipocytes size distribution in eWAT and iWAT and ratio of lipid area in BAT were quantified (n = 6 per group). *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

Article Snippet: Human serum SIRT6, TNFα and TNFR2 concentrations were analysed by Human NAD- dependent deacetylase sirtuin- 6 (SIRT6) ELISA kit (CSB- E17018h, CUSABIO), human tumour necrosis factor α (TNF- α) ELISA kit (CSB- E04740h, CUSABIO) and human soluble tumour necrosis factor receptor 2 (sTNF- R2) ELISA kit (CSB- E11266h, CUSABIO) according to the manufacturer's instructions, individually.

Techniques: Over Expression, Transgenic Assay, Injection, Staining

Journal: Journal of cachexia, sarcopenia and muscle

Article Title: SIRT6 Ameliorates Cancer Cachexia-Associated Adipose Wasting by Suppressing TNFR2 Signalling in Mice.

doi: 10.1002/jcsm.13734

Figure Lengend Snippet: FIGURE 3 | SIRT6 overexpression decreased the expression of genes involved of browning and lipolysis. (A) The mRNA levels of UCP1 in the eWAT were determined by qRT-PCR (n = 4 per group). (B) The mRNA levels of PGC1α in the eWAT were determined by qRT-PCR (n = 4 per group). (C) The expression and phosphorylation levels of lipolysis associated proteins in the eWAT from WT + PBS, WT + LLC, TG + PBS and TG + LLC mice were determined by western blot (n = 6 per group). *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

Article Snippet: Human serum SIRT6, TNFα and TNFR2 concentrations were analysed by Human NAD- dependent deacetylase sirtuin- 6 (SIRT6) ELISA kit (CSB- E17018h, CUSABIO), human tumour necrosis factor α (TNF- α) ELISA kit (CSB- E04740h, CUSABIO) and human soluble tumour necrosis factor receptor 2 (sTNF- R2) ELISA kit (CSB- E11266h, CUSABIO) according to the manufacturer's instructions, individually.

Techniques: Over Expression, Expressing, Quantitative RT-PCR, Phospho-proteomics, Western Blot

Journal: Cell Death & Disease

Article Title: Smooth muscle liver kinase B1 inhibits foam cell formation and atherosclerosis via direct phosphorylation and activation of SIRT6

doi: 10.1038/s41419-023-06054-x

Figure Lengend Snippet: A ChIP-PCR analysis to determine the histone acetylation of LOX-1 promoter in VSMCs infected with adenovirus-expressing GFP or LKB1 ( n = 4). *** P < 0.001 vs GFP. B – D VSMCs were transfected with CTR siRNA or SIRT6 siRNA before infection with adenovirus-expressing GFP or LKB1. B Western blot analysis of LOX-1 protein ( n = 6). * P < 0.05, ** P < 0.01 vs GFP + CTR siRNA, # P < 0.05 vs LKB1 + CTR siRNA. C Oil-red O staining after 24-h stimulation of oxLDL ( n = 6). Scale bar = 10 μm. * P < 0.05 vs GFP + CTR siRNA, # P < 0.05 vs LKB1 + CTR siRNA. D Representative fluorescence images of VSMCs obtained after 4-h incubation with Dil-oxLDL. Scale bar = 10 μm. ** P < 0.01, *** P < 0.001 vs GFP + CTR siRNA, ## P < 0.01 vs LKB1 + CTR siRNA. E – G VSMCs from CTR or LKB1 SMKO mice were transfected with plasmids encoding the murine SIRT6 gene. E Western blot analysis of LOX-1 protein ( n = 6). * P < 0.05, ** P < 0.01 vs CTR+Mock, # P < 0.05 vs LKB1 SMKO +Mock. F Oil-red O staining after 24-h stimulation of oxLDL ( n = 6). Scale bar = 10 μm. * P < 0.05, ** P < 0.01 vs CTR+Mock, # P < 0.05 vs LKB1 SMKO +Mock. G Representative fluorescence images of VSMCs obtained 4-h incubation with Dil-oxLDL ( n = 6). Scale bar = 10 μm. * P < 0.05, *** P < 0.001 vs CTR+Mock, ## P < 0.01 vs LKB1 SMKO +Mock. Data were analyzed by two-tailed Student’s unpaired t -test ( A ) or one-way ANOVA followed by Bonferroni multiple comparison analysis ( B – G ).

Article Snippet: SIRT6 activity was measured using a fluorometric SIRT6 assay kit (ab156068, Abcam), following the manufacturer’s instructions.

Techniques: Infection, Expressing, Transfection, Western Blot, Staining, Fluorescence, Incubation, Two Tailed Test

Journal: Cell Death & Disease

Article Title: Smooth muscle liver kinase B1 inhibits foam cell formation and atherosclerosis via direct phosphorylation and activation of SIRT6

doi: 10.1038/s41419-023-06054-x

Figure Lengend Snippet: A Representative immunofluorescence images of co-localization of LKB1 and SIRT6 in VSMCs. Scale bar = 50 μm. B VSMC lysates were immunoprecipitated with IgG or anti-SIRT6. Results were determined via western blot analysis with anti-LKB1 and anti-SIRT6 antibodies. C, D HEK 293 T cells were transfected with plasmids encoding Flag-tagged LKB1 and HA-tagged SIRT6, and lysates were immunoprecipitated with IgG and anti-Flag ( C ) or anti-HA ( D ) antibodies respectively. Results were determined via western blot analysis with anti-HA and anti-Flag antibodies. E Purified His-LKB1 proteins were incubated with GST or GST-SIRT6-conjugated GSH beads, the boiled eluates were then separated and detected by western blot analysis with anti-His and anti-GST antibodies. F HEK 293 T cells were transfected with plasmids encoding HA-SIRT6-WT/T51A/T57A/T184A and adenovirus expressing GFP or LKB1, lysates were immunoprecipitated with IgG or anti-HA. Results were determined via western blot analysis with anti-P-Ser/Thr and anti-HA antibodies. G , H VSMCs were transfected with plasmids encoding HA-SIRT6-WT/T51A/T57A/T184A. G Western blot analysis of LOX-1 protein ( n = 4). * P < 0.05 vs Mock, # P < 0.05 vs WT. H Representative fluorescence images of VSMCs obtained after 4-h incubation with Dil-oxLDL ( n = 6). Scale bar = 10 μm. * P < 0.05 vs Mock, # P < 0.05, ## P < 0.01 vs WT. I SIRT6 activity measurement of nuclear extract from VSMCs transfected with plasmids encoding HA-SIRT6-WT/T51A/T57A/T184A and LKB1-overexpressing adenovirus or null adenovirus ( n = 4). ** P < 0.01, *** P < 0.001 vs WT+Null, # P < 0.05 vs T51A+Null. J , K VSMCs were transfected with plasmids encoding HA-SIRT6-WT/T57D/T184D. J Western blot analysis of LOX-1 protein ( n = 4). * P < 0.05, *** P < 0.001 vs Mock, # P < 0.05 vs WT. K Representative fluorescence images of VSMCs obtained after 4-h incubation with Dil-oxLDL ( n = 6). Scale bar = 10 μm. * P < 0.05, *** P < 0.001 vs Mock, # P < 0.05 vs WT. Data were analyzed by one-way ANOVA followed by Bonferroni multiple comparison analysis ( G – K ).

Article Snippet: SIRT6 activity was measured using a fluorometric SIRT6 assay kit (ab156068, Abcam), following the manufacturer’s instructions.

Techniques: Immunofluorescence, Immunoprecipitation, Western Blot, Transfection, Purification, Incubation, Expressing, Fluorescence, Activity Assay

Journal: Cell Death & Disease

Article Title: Smooth muscle liver kinase B1 inhibits foam cell formation and atherosclerosis via direct phosphorylation and activation of SIRT6

doi: 10.1038/s41419-023-06054-x

Figure Lengend Snippet: CTR and LKB1 SMKO mice were injected with AAV9-shCON or AAV9-shLOX-1 1 week before injection with rAAV8/D377Y-mPCSK9 and a Paigen diet feeding for 12 weeks. A Oil-red O staining in aortas ( n = 6). * P < 0.05, ** P < 0.01 vs CTR + AAV9-shCON, ## P < 0.01 vs LKB1 SMKO + AAV9-shCON. B Hematoxylin and eosin (H&E) staining in aortic roots ( n = 6). Scale bar = 200 μm. * P < 0.05, *** P < 0.001 vs CTR + AAV9-shCON, ### P < 0.001 vs LKB1 SMKO + AAV9-shCON. C Oil-red O staining in aortic roots ( n =6). Scale bar = 200 μm. * P < 0.05, *** P < 0.001 vs CTR + AAV9-shCON, ### P < 0.001 vs LKB1 SMKO+ AAV9-shCON. D Immunohistochemical staining of MOMA-2 in aortic roots ( n = 6). Scale bar = 200 μm. * P < 0.05, ** P < 0.01 vs CTR + AAV9-shCON, ## P < 0.01 vs LKB1 SMKO + AAV9-shCON. E Double labeling immunofluorescent staining of BODIPY and α-SMA in aortic roots ( n = 6). Scale bar = 100 μm. * P < 0.05, ** P < 0.01 vs CTR + AAV9-shCON, # P < 0.05 vs LKB1 SMKO + AAV9-shCON. F Levels of TC, TG, HDL-C, and LDL-C in serum. Data were analyzed by one-way ANOVA followed by Bonferroni multiple comparison analysis ( A – F ). G Smooth muscle LKB1 inhibits VSMC-derived foam cell formation and atherosclerosis via phosphorylation of SIRT6 and subsequent inhibition of LOX-1 expression.

Article Snippet: SIRT6 activity was measured using a fluorometric SIRT6 assay kit (ab156068, Abcam), following the manufacturer’s instructions.

Techniques: Injection, Staining, Immunohistochemical staining, Labeling, Derivative Assay, Inhibition, Expressing