Review

human sirt2 protein rsirt2 his6 (R&D Systems)

R&D Systems is a verified supplier

R&D Systems manufactures this product

About

News

Press Release

Team

Advisors

Partners

Contact

Bioz Stars

Bioz vStars

Buy from Supplier

Structured Review

R&D Systems human sirt2 protein rsirt2 his6

Figure 1. (a) Mass spectrometry was used to quantify acetylated peptides from male <t>Sirt2+/+</t> (WT) and <t>Sirt2−/−(KO)</t> livers. Peptides with significant changes in acetylation are rendered in red. Blue lines indicate the mean log2 fold-change. A total of 2452 acetylated peptides were quantified. The mean acetylation level significantly increased in 317 peptides (p < 0.01, FC > 1.5) due to Sirt2 ablation. (b) Mass spectrometry was used to quantify acetylated peptides from female Sirt2+/+ and Sirt2−/− livers. There was no appreciable change in acetylation levels across putative Sirt2-targets. (c) Volcano plot of Sirt2+/+ female to male ratios at each of the 2452 putative Sirt2 targets, expressed as Log2 FC. (d) Volcano plot of Sirt2−/−female to male ratios at each of the 2452 putative Sirt2 targets, expressed as Log2 FC. (e) The mean Log2 FC in Sirt2+/+ female/male acetylation was 4.2; the mean Log2 FC in Sirt2−/−female/male acetylation was 1.3. Quantification and comparison of the Log2 FC ratios using a Student’s two-sided t-test showed that Sirt2−/−males had significant decrease in hyperacetylation in 2452 putative sites compared to wild type counterparts (p < 0.0001). (f) Immunoblotting for Sirt2 in livers from wild type mice indicated no sex difference in the expression of the Sirt2 protein. Note that there are multiple bands due to multiple isoforms of Sirt2. **** p < 0.0001 by Student’s two-sided t-test.

Human Sirt2 Protein Rsirt2 His6, supplied by R&D Systems, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/recombinant+human+sirt2/Recombinant+Human+Sirtuin+2%2FSIRT2+Protein%2C+CF/pm39334926-61-26-30
Average 94 stars, based on 1 article reviews

human sirt2 protein rsirt2 his6 - by Bioz Stars, 2026-10

94/100 stars

Images

1) Product Images from "Sirt2 Regulates Liver Metabolism in a Sex-Specific Manner."

Article Title: Sirt2 Regulates Liver Metabolism in a Sex-Specific Manner.

Journal: Biomolecules

doi: 10.3390/biom14091160

Figure 1. (a) Mass spectrometry was used to quantify acetylated peptides from male Sirt2+/+ (WT) and Sirt2−/−(KO) livers. Peptides with significant changes in acetylation are rendered in red. Blue lines indicate the mean log2 fold-change. A total of 2452 acetylated peptides were quantified. The mean acetylation level significantly increased in 317 peptides (p < 0.01, FC > 1.5) due to Sirt2 ablation. (b) Mass spectrometry was used to quantify acetylated peptides from female Sirt2+/+ and Sirt2−/− livers. There was no appreciable change in acetylation levels across putative Sirt2-targets. (c) Volcano plot of Sirt2+/+ female to male ratios at each of the 2452 putative Sirt2 targets, expressed as Log2 FC. (d) Volcano plot of Sirt2−/−female to male ratios at each of the 2452 putative Sirt2 targets, expressed as Log2 FC. (e) The mean Log2 FC in Sirt2+/+ female/male acetylation was 4.2; the mean Log2 FC in Sirt2−/−female/male acetylation was 1.3. Quantification and comparison of the Log2 FC ratios using a Student’s two-sided t-test showed that Sirt2−/−males had significant decrease in hyperacetylation in 2452 putative sites compared to wild type counterparts (p < 0.0001). (f) Immunoblotting for Sirt2 in livers from wild type mice indicated no sex difference in the expression of the Sirt2 protein. Note that there are multiple bands due to multiple isoforms of Sirt2. **** p < 0.0001 by Student’s two-sided t-test.

Figure Legend Snippet: Figure 1. (a) Mass spectrometry was used to quantify acetylated peptides from male Sirt2+/+ (WT) and Sirt2−/−(KO) livers. Peptides with significant changes in acetylation are rendered in red. Blue lines indicate the mean log2 fold-change. A total of 2452 acetylated peptides were quantified. The mean acetylation level significantly increased in 317 peptides (p < 0.01, FC > 1.5) due to Sirt2 ablation. (b) Mass spectrometry was used to quantify acetylated peptides from female Sirt2+/+ and Sirt2−/− livers. There was no appreciable change in acetylation levels across putative Sirt2-targets. (c) Volcano plot of Sirt2+/+ female to male ratios at each of the 2452 putative Sirt2 targets, expressed as Log2 FC. (d) Volcano plot of Sirt2−/−female to male ratios at each of the 2452 putative Sirt2 targets, expressed as Log2 FC. (e) The mean Log2 FC in Sirt2+/+ female/male acetylation was 4.2; the mean Log2 FC in Sirt2−/−female/male acetylation was 1.3. Quantification and comparison of the Log2 FC ratios using a Student’s two-sided t-test showed that Sirt2−/−males had significant decrease in hyperacetylation in 2452 putative sites compared to wild type counterparts (p < 0.0001). (f) Immunoblotting for Sirt2 in livers from wild type mice indicated no sex difference in the expression of the Sirt2 protein. Note that there are multiple bands due to multiple isoforms of Sirt2. **** p < 0.0001 by Student’s two-sided t-test.

Techniques Used: Mass Spectrometry, Comparison, Western Blot, Expressing

$Figure 2. (a) The 306 putative Sirt2 target sites mapped to proteins spread across multiple cellular compartments but were most abundant in the mitochondria and cytosol. (b) A plot of the Log2 ratio (FC > 1.5) of Sirt2−/−vs. Sirt2+/+ by cellular compartment revealed that the mitochondria and cytosol had the largest FCs in lysine acetylation. (c) The Log2 Ratio of putative mitochondrial Sirt2 targets from this study was compared to known Sirt3 targets within the mitochondria. Of the 140 Sirt2 targets represented in this plot, 19% are also Sirt3 targets. (d) Preparations of Sirt2+/+ nuclear and cytosolic fractions show that Sirt2 is present in both compartments, confirmed with the cytosolic marker α-tubulin and the nuclear marker, Lamin A/C (1 and 2 denote matched, replicate samples). Sirt2 is present in both compartments, confirming acetylomics data. (e) Sirt2 was not detected in mitochondria prepared from Sirt2+/+ hepatic tissue lysates (N = 3). 1–3 denotes replicate samples. (f) Sirt2 was not detected in peroxisomes prepared from Sirt2+/+ hepatic tissue lysates (N = 3). 1–3 denotes replicate samples. (g) The ultrapure subcellular fractionation technique used completely separated mitochondria and peroxisomes, confirmed with the peroxisomal marker, ACOX1, and the mitochondrial marker, TIMM23 (N = 3). 1–3 denotes matched, replicate samples. Membranes exposed at the same time, cropped for clarity.$
Figure Legend Snippet: Figure 2. (a) The 306 putative Sirt2 target sites mapped to proteins spread across multiple cellular compartments but were most abundant in the mitochondria and cytosol. (b) A plot of the Log2 ratio (FC > 1.5) of Sirt2−/−vs. Sirt2+/+ by cellular compartment revealed that the mitochondria and cytosol had the largest FCs in lysine acetylation. (c) The Log2 Ratio of putative mitochondrial Sirt2 targets from this study was compared to known Sirt3 targets within the mitochondria. Of the 140 Sirt2 targets represented in this plot, 19% are also Sirt3 targets. (d) Preparations of Sirt2+/+ nuclear and cytosolic fractions show that Sirt2 is present in both compartments, confirmed with the cytosolic marker α-tubulin and the nuclear marker, Lamin A/C (1 and 2 denote matched, replicate samples). Sirt2 is present in both compartments, confirming acetylomics data. (e) Sirt2 was not detected in mitochondria prepared from Sirt2+/+ hepatic tissue lysates (N = 3). 1–3 denotes replicate samples. (f) Sirt2 was not detected in peroxisomes prepared from Sirt2+/+ hepatic tissue lysates (N = 3). 1–3 denotes replicate samples. (g) The ultrapure subcellular fractionation technique used completely separated mitochondria and peroxisomes, confirmed with the peroxisomal marker, ACOX1, and the mitochondrial marker, TIMM23 (N = 3). 1–3 denotes matched, replicate samples. Membranes exposed at the same time, cropped for clarity.

Techniques Used: Marker, Fractionation

Figure 3. (a,b) Total body weight and lean body mass were the same between Sirt2−/−(N = 11) and Sirt2+/+ (N = 8) males, but Sirt2−/−males had significantly lower fat mass compared to Sirt2+/+ males (p < 0.05). (b) Total body weight, fat mass, and lean mass were not significantly different between Sirt2−/−(N = 6) and Sirt2+/+ females (N = 8). Body composition was calculated for each mouse by dividing fat mass by lean mass. Only Sirt2−/−males had a significantly reduced fat/lean mass body composition (p < 0.001) compared to controls; female Sirt2−/−did not. (c) Blood glucose levels in Sirt2−/−vs. Sirt2+/+ males or females in both a fed and fasted state. In a fed state, there was no difference in blood glucose levels between Sirt2−/−and Sirt2+/+ in either males (N = 8) or females (N = 8). Upon fasting, only Sirt2−/−males had significantly reduced blood glucose levels (N = 8; p < 0.05) while Sirt2−/−females showed no significant reduction compared to controls (N = 8). (d) H&E staining of Sirt2−/−vs. Sirt2+/+ livers indicated no obvious morphological changes or tissue damage (N = 4). (e) Male Sirt2−/−mice (N = 3) had significantly less triglycerides in hepatic tissue compared to Sirt2+/+ males (N = 3; p < 0.05). Comparisons made with a Student’s nonparametric t-test; * p < 0.05, *** p < 0.001.

Figure Legend Snippet: Figure 3. (a,b) Total body weight and lean body mass were the same between Sirt2−/−(N = 11) and Sirt2+/+ (N = 8) males, but Sirt2−/−males had significantly lower fat mass compared to Sirt2+/+ males (p < 0.05). (b) Total body weight, fat mass, and lean mass were not significantly different between Sirt2−/−(N = 6) and Sirt2+/+ females (N = 8). Body composition was calculated for each mouse by dividing fat mass by lean mass. Only Sirt2−/−males had a significantly reduced fat/lean mass body composition (p < 0.001) compared to controls; female Sirt2−/−did not. (c) Blood glucose levels in Sirt2−/−vs. Sirt2+/+ males or females in both a fed and fasted state. In a fed state, there was no difference in blood glucose levels between Sirt2−/−and Sirt2+/+ in either males (N = 8) or females (N = 8). Upon fasting, only Sirt2−/−males had significantly reduced blood glucose levels (N = 8; p < 0.05) while Sirt2−/−females showed no significant reduction compared to controls (N = 8). (d) H&E staining of Sirt2−/−vs. Sirt2+/+ livers indicated no obvious morphological changes or tissue damage (N = 4). (e) Male Sirt2−/−mice (N = 3) had significantly less triglycerides in hepatic tissue compared to Sirt2+/+ males (N = 3; p < 0.05). Comparisons made with a Student’s nonparametric t-test; * p < 0.05, *** p < 0.001.

Techniques Used: Staining

Figure 4. (a) Sirt2−/−and Sirt2+/+ males ran the same distance (N = 5) before reaching exhaustion. (b) Blood glucose levels were normal prior to the treadmill assay in male Sirt2−/−and Sirt2+/+ mice. Upon exhaustion, male Sirt2−/−mice (N = 5) had significantly lower blood glucose levels than Sirt2+/+ counterparts (N = 5; p < 0.01). (c) Sirt2−/−females (N = 6) ran significantly further than Sirt2+/+ females (N = 7; p < 0.005) before reaching exhaustion. (d) There was no significant difference in blood glucose levels between Sirt2−/−(N = 6) and Sirt2+/+ females (N = 7) post-exhaustion. Comparisons were made using a Student’s nonparametric t-test; ns = no difference; ** p < 0.01.

Figure Legend Snippet: Figure 4. (a) Sirt2−/−and Sirt2+/+ males ran the same distance (N = 5) before reaching exhaustion. (b) Blood glucose levels were normal prior to the treadmill assay in male Sirt2−/−and Sirt2+/+ mice. Upon exhaustion, male Sirt2−/−mice (N = 5) had significantly lower blood glucose levels than Sirt2+/+ counterparts (N = 5; p < 0.01). (c) Sirt2−/−females (N = 6) ran significantly further than Sirt2+/+ females (N = 7; p < 0.005) before reaching exhaustion. (d) There was no significant difference in blood glucose levels between Sirt2−/−(N = 6) and Sirt2+/+ females (N = 7) post-exhaustion. Comparisons were made using a Student’s nonparametric t-test; ns = no difference; ** p < 0.01.

Techniques Used:

Figure 5. (a) Male Sirt2−/−mice had a blunted response to a bolus lactate injection, with significantly decreased glucose production 15-min post-injection (N = 5; p < 0.05). (b) Sirt2−/−females had a

Figure Legend Snippet: Figure 5. (a) Male Sirt2−/−mice had a blunted response to a bolus lactate injection, with significantly decreased glucose production 15-min post-injection (N = 5; p < 0.05). (b) Sirt2−/−females had a

Techniques Used: Injection

Figure 6. Schematic of glycolytic enzymes (red) and gluconeogenic enzymes (purple) that contain putative Sirt2 targets. Each significantly altered residue (p < 0.01; FC > 1.5) is identified with FC reported. Non-targets are in blue.

Figure Legend Snippet: Figure 6. Schematic of glycolytic enzymes (red) and gluconeogenic enzymes (purple) that contain putative Sirt2 targets. Each significantly altered residue (p < 0.01; FC > 1.5) is identified with FC reported. Non-targets are in blue.

Techniques Used: Residue

Figure 7. (a) Hepatocytes isolated from a Sirt2−/−liver showed reduced OCR compared to Sirt2+/+ hepatocytes during a mitochondrial stress test using a Seahorse Extracellular Flux Analyzer (p < 0.005). (b) Hepatocytes isolated from Sirt2−/−liver showed significantly reduced OCR compared to Sirt2+/+

Figure Legend Snippet: Figure 7. (a) Hepatocytes isolated from a Sirt2−/−liver showed reduced OCR compared to Sirt2+/+ hepatocytes during a mitochondrial stress test using a Seahorse Extracellular Flux Analyzer (p < 0.005). (b) Hepatocytes isolated from Sirt2−/−liver showed significantly reduced OCR compared to Sirt2+/+

Techniques Used: Isolation

Recombinant:

Article Title: Facile chemoenzymatic synthesis of a novel stable mimic of NAD + †The X-ray structure of S-NAD + with human CD38 is deposited in Protein Data Bank ( https://www.rcsb.org ) (PDB ID: 6EDR ). ‡Electronic supplementary information (ESI) available: Experimental procedures, and supplementary figures, table, and data. See DOI: 10.1039/c8sc03899f
Article Snippet: HPLC analysis was performed with a semipreparative C18 Kinetex column (5 μm, 100 Å, 150×10.0 mm, from Phenomenex Inc, Torrance, CA) using following method: mobile phase A: 0.1% formic acid (aq); mobile phase B: 0.1% formic acid in methanol; flow rate: 2.0 ml/min; 0-2 min: 0-4% B, 2-4 min: 4-10% B; 4-6 min: 10-20% B; 6-9 min: 20% B; 9-12 min: 20-50% B; and 12-14 min: 50-0% B. Substrate activity of S-NAD+ for human sirtuins 2 (SIRT2). .. Recombinant human SIRT2 and a fluorogenic peptide substrate Ac-RGK(Ac)- AMC were purchased from R&D Systems (Minneapolis, MN). .. Bovine trypsin was purchased from Sigma-Aldrich (St. Louis, MO).

Article Title: A ribose-functionalized NAD + with unexpected high activity and selectivity for protein poly-ADP-ribosylation
Article Snippet: .. 0.5 mM NAD + or 6 was incubated with 0.25 mM acetylated peptide substrate Ac-Arg-Gly-Lys(Ac)-AMC (R&D System, MN) and 45.4 nM recombinant human SIRT2 (R&D System, MN) in buffer containing 25 mM Tris pH 8.0, 150 mM NaCl, 1 mM DTT for overnight at room temperature. ..

Article Title: A ribose-functionalized NAD + with unexpected high activity and selectivity for protein poly-ADP-ribosylation.
Article Snippet: .. 0.5mM NAD+ or 6 was incubated with 0.25 mM acetylated peptide substrate Ac-Arg-Gly-Lys(Ac)-AMC (R&D System, MN) and 45.4 nM recombinant human SIRT2 (R&D System, MN) in buffer containing 25mM Tris pH 8.0, 150mM NaCl, 1 mM DTT for overnight at room temperature. ..

Incubation:

Image Search Results

Journal: Biomolecules

Article Title: Sirt2 Regulates Liver Metabolism in a Sex-Specific Manner.

doi: 10.3390/biom14091160

Figure Lengend Snippet: Figure 1. (a) Mass spectrometry was used to quantify acetylated peptides from male Sirt2+/+ (WT) and Sirt2−/−(KO) livers. Peptides with significant changes in acetylation are rendered in red. Blue lines indicate the mean log2 fold-change. A total of 2452 acetylated peptides were quantified. The mean acetylation level significantly increased in 317 peptides (p < 0.01, FC > 1.5) due to Sirt2 ablation. (b) Mass spectrometry was used to quantify acetylated peptides from female Sirt2+/+ and Sirt2−/− livers. There was no appreciable change in acetylation levels across putative Sirt2-targets. (c) Volcano plot of Sirt2+/+ female to male ratios at each of the 2452 putative Sirt2 targets, expressed as Log2 FC. (d) Volcano plot of Sirt2−/−female to male ratios at each of the 2452 putative Sirt2 targets, expressed as Log2 FC. (e) The mean Log2 FC in Sirt2+/+ female/male acetylation was 4.2; the mean Log2 FC in Sirt2−/−female/male acetylation was 1.3. Quantification and comparison of the Log2 FC ratios using a Student’s two-sided t-test showed that Sirt2−/−males had significant decrease in hyperacetylation in 2452 putative sites compared to wild type counterparts (p < 0.0001). (f) Immunoblotting for Sirt2 in livers from wild type mice indicated no sex difference in the expression of the Sirt2 protein. Note that there are multiple bands due to multiple isoforms of Sirt2. **** p < 0.0001 by Student’s two-sided t-test.

Article Snippet: A total of 1 mg of liver homogenate was pre-cleared with anti-His6 (1:500, Roche, St. Louis, MO, USA) then incubated with 20 μg of recombinant His6-tagged human SIRT2 protein (rSIRT2-His6) (R&D Systems, Minneapolis, MN, USA) at 37 ◦C for 30 min in a microfuge rotator.

Techniques: Mass Spectrometry, Comparison, Western Blot, Expressing

Journal: Biomolecules

Article Title: Sirt2 Regulates Liver Metabolism in a Sex-Specific Manner.

doi: 10.3390/biom14091160

Figure Lengend Snippet: Figure 2. (a) The 306 putative Sirt2 target sites mapped to proteins spread across multiple cellular compartments but were most abundant in the mitochondria and cytosol. (b) A plot of the Log2 ratio (FC > 1.5) of Sirt2−/−vs. Sirt2+/+ by cellular compartment revealed that the mitochondria and cytosol had the largest FCs in lysine acetylation. (c) The Log2 Ratio of putative mitochondrial Sirt2 targets from this study was compared to known Sirt3 targets within the mitochondria. Of the 140 Sirt2 targets represented in this plot, 19% are also Sirt3 targets. (d) Preparations of Sirt2+/+ nuclear and cytosolic fractions show that Sirt2 is present in both compartments, confirmed with the cytosolic marker α-tubulin and the nuclear marker, Lamin A/C (1 and 2 denote matched, replicate samples). Sirt2 is present in both compartments, confirming acetylomics data. (e) Sirt2 was not detected in mitochondria prepared from Sirt2+/+ hepatic tissue lysates (N = 3). 1–3 denotes replicate samples. (f) Sirt2 was not detected in peroxisomes prepared from Sirt2+/+ hepatic tissue lysates (N = 3). 1–3 denotes replicate samples. (g) The ultrapure subcellular fractionation technique used completely separated mitochondria and peroxisomes, confirmed with the peroxisomal marker, ACOX1, and the mitochondrial marker, TIMM23 (N = 3). 1–3 denotes matched, replicate samples. Membranes exposed at the same time, cropped for clarity.

Techniques: Marker, Fractionation

Journal: Biomolecules

Article Title: Sirt2 Regulates Liver Metabolism in a Sex-Specific Manner.

doi: 10.3390/biom14091160

Figure Lengend Snippet: Figure 3. (a,b) Total body weight and lean body mass were the same between Sirt2−/−(N = 11) and Sirt2+/+ (N = 8) males, but Sirt2−/−males had significantly lower fat mass compared to Sirt2+/+ males (p < 0.05). (b) Total body weight, fat mass, and lean mass were not significantly different between Sirt2−/−(N = 6) and Sirt2+/+ females (N = 8). Body composition was calculated for each mouse by dividing fat mass by lean mass. Only Sirt2−/−males had a significantly reduced fat/lean mass body composition (p < 0.001) compared to controls; female Sirt2−/−did not. (c) Blood glucose levels in Sirt2−/−vs. Sirt2+/+ males or females in both a fed and fasted state. In a fed state, there was no difference in blood glucose levels between Sirt2−/−and Sirt2+/+ in either males (N = 8) or females (N = 8). Upon fasting, only Sirt2−/−males had significantly reduced blood glucose levels (N = 8; p < 0.05) while Sirt2−/−females showed no significant reduction compared to controls (N = 8). (d) H&E staining of Sirt2−/−vs. Sirt2+/+ livers indicated no obvious morphological changes or tissue damage (N = 4). (e) Male Sirt2−/−mice (N = 3) had significantly less triglycerides in hepatic tissue compared to Sirt2+/+ males (N = 3; p < 0.05). Comparisons made with a Student’s nonparametric t-test; * p < 0.05, *** p < 0.001.

Techniques: Staining

Journal: Biomolecules

Article Title: Sirt2 Regulates Liver Metabolism in a Sex-Specific Manner.

doi: 10.3390/biom14091160

Figure Lengend Snippet: Figure 4. (a) Sirt2−/−and Sirt2+/+ males ran the same distance (N = 5) before reaching exhaustion. (b) Blood glucose levels were normal prior to the treadmill assay in male Sirt2−/−and Sirt2+/+ mice. Upon exhaustion, male Sirt2−/−mice (N = 5) had significantly lower blood glucose levels than Sirt2+/+ counterparts (N = 5; p < 0.01). (c) Sirt2−/−females (N = 6) ran significantly further than Sirt2+/+ females (N = 7; p < 0.005) before reaching exhaustion. (d) There was no significant difference in blood glucose levels between Sirt2−/−(N = 6) and Sirt2+/+ females (N = 7) post-exhaustion. Comparisons were made using a Student’s nonparametric t-test; ns = no difference; ** p < 0.01.

Techniques:

Journal: Biomolecules

Article Title: Sirt2 Regulates Liver Metabolism in a Sex-Specific Manner.

doi: 10.3390/biom14091160

Figure Lengend Snippet: Figure 5. (a) Male Sirt2−/−mice had a blunted response to a bolus lactate injection, with significantly decreased glucose production 15-min post-injection (N = 5; p < 0.05). (b) Sirt2−/−females had a

Techniques: Injection

Journal: Biomolecules

Article Title: Sirt2 Regulates Liver Metabolism in a Sex-Specific Manner.

doi: 10.3390/biom14091160

Figure Lengend Snippet: Figure 6. Schematic of glycolytic enzymes (red) and gluconeogenic enzymes (purple) that contain putative Sirt2 targets. Each significantly altered residue (p < 0.01; FC > 1.5) is identified with FC reported. Non-targets are in blue.

Techniques: Residue

Journal: Biomolecules

Article Title: Sirt2 Regulates Liver Metabolism in a Sex-Specific Manner.

doi: 10.3390/biom14091160

Figure Lengend Snippet: Figure 7. (a) Hepatocytes isolated from a Sirt2−/−liver showed reduced OCR compared to Sirt2+/+ hepatocytes during a mitochondrial stress test using a Seahorse Extracellular Flux Analyzer (p < 0.005). (b) Hepatocytes isolated from Sirt2−/−liver showed significantly reduced OCR compared to Sirt2+/+

Techniques: Isolation

a , b TNF-α levels of (a) BMDMs and (b) J774A.1 macrophages primed with LPS and then treated with rhein or DMSO for 0.5 h and finally stimulated with ATP. c Immunoblots of phosphorylation, ubiquitylation, acetylation, and sumoylation levels in the cell lysate of BMDMs and J774A.1 macrophages treated as ( a , b ). d The workflow of acetylome analysis. e The Venn diagram of 152 proteins with a significant change of acetylation after rhein treatment. f Heat map of 25 NLRP3 inflammasome-related proteins among 152 proteins in ( e ). g Counts on connections of 25 proteins in f to NLRP3-related cytoplasmic acetylation modifying enzymes. h – j J774A.1 macrophages transfected with si- Sirt2 or si- Scramble were primed with LPS and then treated with rhein or DMSO for 0.5 h and finally stimulated with ATP. h PI-positive cells were detected by flowcytometry. i Immunoblots of Caspase1, SIRT2 and acetylated α-Tubulin in cell lysate. j Percentage of PI-positive cells in ( h ). Data are presented as the mean ± SD. Significance is indicated by ** p < 0.01 compared to the corresponding group. n.s. indicates no significance.

Journal: Communications Biology

Article Title: Rhein targets macrophage SIRT2 to promote adipose tissue thermogenesis in obesity in mice

doi: 10.1038/s42003-024-06693-6

Figure Lengend Snippet: a , b TNF-α levels of (a) BMDMs and (b) J774A.1 macrophages primed with LPS and then treated with rhein or DMSO for 0.5 h and finally stimulated with ATP. c Immunoblots of phosphorylation, ubiquitylation, acetylation, and sumoylation levels in the cell lysate of BMDMs and J774A.1 macrophages treated as ( a , b ). d The workflow of acetylome analysis. e The Venn diagram of 152 proteins with a significant change of acetylation after rhein treatment. f Heat map of 25 NLRP3 inflammasome-related proteins among 152 proteins in ( e ). g Counts on connections of 25 proteins in f to NLRP3-related cytoplasmic acetylation modifying enzymes. h – j J774A.1 macrophages transfected with si- Sirt2 or si- Scramble were primed with LPS and then treated with rhein or DMSO for 0.5 h and finally stimulated with ATP. h PI-positive cells were detected by flowcytometry. i Immunoblots of Caspase1, SIRT2 and acetylated α-Tubulin in cell lysate. j Percentage of PI-positive cells in ( h ). Data are presented as the mean ± SD. Significance is indicated by ** p < 0.01 compared to the corresponding group. n.s. indicates no significance.

Article Snippet: Briefly, the sensor slide was activated by carbodiimide and N-hydroxysuccinimide, and human recombinant SIRT2 protein (CaymanChemical, US) was diluted in acetate solution pH 4.0 and immobilized on the surface of one channel of the sensor with a neighboring channel serving as a reference.

Techniques: Western Blot, Phospho-proteomics, Transfection

a , b BMDMs were primed with LPS and pretreated with SIRT2 inhibitor AGK2 for 2 h, and then treated with rhein or DMSO for 0.5 h and finally stimulated with ATP. a IL-1β levels detected by ELISA in the supernatant of BMDMs. b Immunoblots of Caspase1 and acetylated α-Tubulin in cell lysate of BMDMs. c The ratio of NAD + to NADH in BMDMs and J774A.1 macrophages primed with LPS and then treated with rhein or DMSO for 0.5 h and finally stimulated with ATP. d SPR assays on recombinant mouse SIRT2 incubated with rhein at the indicated concentrations. e MST assays on cell lysate of HEK-293T cells transfected with SIRT2-GFP incubated with rhein at the indicated concentrations. f Merge of the catalytic domain and highly flexible region of SIRT2 captured molecular dynamics simulations. g Keyframe of rhein-SIRT2 combination captured in molecular dynamics simulations. The dashed lines represent hydrogen bonds. Data are presented as the mean ± SD. Significance is indicated by * p < 0.05 and *** p < 0.001 compared to the corresponding group. n.s. indicates no significance.

Journal: Communications Biology

Article Title: Rhein targets macrophage SIRT2 to promote adipose tissue thermogenesis in obesity in mice

doi: 10.1038/s42003-024-06693-6

Figure Lengend Snippet: a , b BMDMs were primed with LPS and pretreated with SIRT2 inhibitor AGK2 for 2 h, and then treated with rhein or DMSO for 0.5 h and finally stimulated with ATP. a IL-1β levels detected by ELISA in the supernatant of BMDMs. b Immunoblots of Caspase1 and acetylated α-Tubulin in cell lysate of BMDMs. c The ratio of NAD + to NADH in BMDMs and J774A.1 macrophages primed with LPS and then treated with rhein or DMSO for 0.5 h and finally stimulated with ATP. d SPR assays on recombinant mouse SIRT2 incubated with rhein at the indicated concentrations. e MST assays on cell lysate of HEK-293T cells transfected with SIRT2-GFP incubated with rhein at the indicated concentrations. f Merge of the catalytic domain and highly flexible region of SIRT2 captured molecular dynamics simulations. g Keyframe of rhein-SIRT2 combination captured in molecular dynamics simulations. The dashed lines represent hydrogen bonds. Data are presented as the mean ± SD. Significance is indicated by * p < 0.05 and *** p < 0.001 compared to the corresponding group. n.s. indicates no significance.

Techniques: Enzyme-linked Immunosorbent Assay, Western Blot, Recombinant, Incubation, Transfection

a Workflow for the administration of rhein in myeloid Sirt2 -knockdown mice. b Daily body weight of control and myeloid Sirt2 -knockdown mice during rhein treatment ( n = 4). c Oral glucose tolerance test on control and Sirt2 -knockdown mice after a 12-h fast ( n = 4). d i.p. insulin tolerance on control and Sirt2 -knockdown mice after 6-h fast ( n = 4). e , f Immunoblots of UCP1 in e iWAT and f eWAT in myeloid Sirt2 -knockdown mice after 270-min cold exposure. Data are presented as the mean ± SD. Significance is indicated by * p < 0.05, ** p < 0.01, and *** p < 0.001 compared to the corresponding group. n.s. indicates no significance.

Journal: Communications Biology

Article Title: Rhein targets macrophage SIRT2 to promote adipose tissue thermogenesis in obesity in mice

doi: 10.1038/s42003-024-06693-6

Figure Lengend Snippet: a Workflow for the administration of rhein in myeloid Sirt2 -knockdown mice. b Daily body weight of control and myeloid Sirt2 -knockdown mice during rhein treatment ( n = 4). c Oral glucose tolerance test on control and Sirt2 -knockdown mice after a 12-h fast ( n = 4). d i.p. insulin tolerance on control and Sirt2 -knockdown mice after 6-h fast ( n = 4). e , f Immunoblots of UCP1 in e iWAT and f eWAT in myeloid Sirt2 -knockdown mice after 270-min cold exposure. Data are presented as the mean ± SD. Significance is indicated by * p < 0.05, ** p < 0.01, and *** p < 0.001 compared to the corresponding group. n.s. indicates no significance.

Techniques: Knockdown, Control, Western Blot

During obesity, rhein promotes adipose tissue thermogenesis by suppressing the activation of NLRP3 inflammasome in adipose tissue-resident macrophages. Furthermore, SIRT2 is a direct molecular target of rhein, and rhein inhibits the activation of NLRP3 inflammasome and promotes adipose tissue thermogenesis in a macrophage SIRT2-dependent manner.

Journal: Communications Biology

Article Title: Rhein targets macrophage SIRT2 to promote adipose tissue thermogenesis in obesity in mice

doi: 10.1038/s42003-024-06693-6

Figure Lengend Snippet: During obesity, rhein promotes adipose tissue thermogenesis by suppressing the activation of NLRP3 inflammasome in adipose tissue-resident macrophages. Furthermore, SIRT2 is a direct molecular target of rhein, and rhein inhibits the activation of NLRP3 inflammasome and promotes adipose tissue thermogenesis in a macrophage SIRT2-dependent manner.

Techniques: Activation Assay

Review

human sirt2 protein rsirt2 his6 (R&D Systems)

Structured Review

Images

1) Product Images from "Sirt2 Regulates Liver Metabolism in a Sex-Specific Manner."

Related Articles

Recombinant:

Incubation:

Similar Products

Image Search Results