Journal: Neurobiology of Stress

Article Title: Microglial SIRT6 confers protection against neuroinflammation-associated depression through NRF2-HO1 signaling

doi: 10.1016/j.ynstr.2026.100804

Figure Lengend Snippet: Hippocampal SIRT6 Expression Profile in a Lipopolysaccharide (LPS)-Induced Depression Model. (A) Western blot analysis of SIRT6 expression in the hippocampal brain tissues. D, day. n = 4 mice. (B) Immunostaining for SIRT6 in the hippocampal region three days post-LPS injection revealed an overall reduction in brain tissues, while its expression was concurrently increased specifically in microglia. This was quantified in two graphs showing total SIRT6 expression (left) and microglial SIRT6 expression (right), with arrowheads indicating SIRT6 localization in microglia. n = 4 mice. (C) Western blot analysis of SIRT6 expression in microglia sorted from the hippocampus by MCAS at 3 days post-LPS injection. n = 4 mice. Data are presented as mean ± SEM. Statistical analysis: One-way ANOVA with Tukey's post hoc test in A; Unpaired t -test for two-group comparisons in B-C.

Article Snippet: Membranes were blocked with 5% non-fat milk in TBST for 1 h and incubated overnight at 4 °C with primary antibodies: SIRT6 (12486, Cell Signaling Technology), NRF2 (ab62352, Abcam), HO-1 (ab68477, Abcam), NQO1 (ab80588, Abcam), KEAP1 (8047, Cell Signaling Technology), NLRP3 (AG-20B-0014, AdipoGen), cleaved caspase-1 (4199, Cell Signaling Technology), cleaved IL-1β (83186, Cell Signaling Technology), and β-actin (A1978, Sigma-Aldrich).

Techniques: Expressing, Western Blot, Immunostaining, Injection

Journal: Neurobiology of Stress

Article Title: Microglial SIRT6 confers protection against neuroinflammation-associated depression through NRF2-HO1 signaling

doi: 10.1016/j.ynstr.2026.100804

Figure Lengend Snippet: Microglial-specific SIRT6 deficiency exacerbates LPS-induced depressive-like behaviors. (A) Experimental timeline showing the administration of tamoxifen to Sirt6 MCKO mice, followed by subsequent LPS injection and behavioral testing. (B) Immunostaining for SIRT6 verified successful microglial SIRT6 knockout in Sirt6 MCKO mice. (C) Western blot analysis verified successful microglial SIRT6 knockout in microglial. n = 3 mice. (D) Open field test (OFT) analysis. Representative trajectories are shown, showcasing the movement patterns of Sirt6 MCKO and Sirt6 fl/fl mice. Data are presented as the time spent in the central area and the total distance traveled. n = 8 mice. (E) Elevated plus maze test (EPM). Representative exploration paths are shown. Data are presented as the number of entries into the open arms and the time spent in the open arms. n = 8 mice. (F) Schematic diagram of the tail suspension test (TST). The immobility time during the test was quantified for each group. n = 8 mice. (G) Schematic diagram of the forced swim test (FST). The immobility time during the test was quantified. n = 8 mice. Data are presented as mean ± SEM. Statistical significance between two groups was determined by an unpaired two-tailed Student's t-test.

Article Snippet: Membranes were blocked with 5% non-fat milk in TBST for 1 h and incubated overnight at 4 °C with primary antibodies: SIRT6 (12486, Cell Signaling Technology), NRF2 (ab62352, Abcam), HO-1 (ab68477, Abcam), NQO1 (ab80588, Abcam), KEAP1 (8047, Cell Signaling Technology), NLRP3 (AG-20B-0014, AdipoGen), cleaved caspase-1 (4199, Cell Signaling Technology), cleaved IL-1β (83186, Cell Signaling Technology), and β-actin (A1978, Sigma-Aldrich).

Techniques: Injection, Immunostaining, Knock-Out, Western Blot, Suspension, Two Tailed Test

Journal: Neurobiology of Stress

Article Title: Microglial SIRT6 confers protection against neuroinflammation-associated depression through NRF2-HO1 signaling

doi: 10.1016/j.ynstr.2026.100804

Figure Lengend Snippet: SIRT6 deficiency potentiates microglial activation in a LPS-induced depression model. (A) Immunostaining of GFAP and IBA1 in the hippocampus area in Sirt6 MCKO and Sirt6 fl/fl mice at day 5 post-LPS injection. n = 4 mice. (B-G) Quantification of (B) IBA1 + and (C) GFAP + cell counts, along with microglial morphology parameters: (D) number of branches, (E) average branch length, (F) total branch length, and (G) soma area. n = 4 mice. (H) qPCR analysis of TNF-α,IL-6, and IL-1β mRNA levels in in the microglial sorted from the hippocampus brain tissue. n = 4 mice. Data are mean ± SEM. Statistical significance between two groups was determined by an unpaired two-tailed Student's t-test.

Article Snippet: Membranes were blocked with 5% non-fat milk in TBST for 1 h and incubated overnight at 4 °C with primary antibodies: SIRT6 (12486, Cell Signaling Technology), NRF2 (ab62352, Abcam), HO-1 (ab68477, Abcam), NQO1 (ab80588, Abcam), KEAP1 (8047, Cell Signaling Technology), NLRP3 (AG-20B-0014, AdipoGen), cleaved caspase-1 (4199, Cell Signaling Technology), cleaved IL-1β (83186, Cell Signaling Technology), and β-actin (A1978, Sigma-Aldrich).

Techniques: Activation Assay, Immunostaining, Injection, Two Tailed Test

Journal: Neurobiology of Stress

Article Title: Microglial SIRT6 confers protection against neuroinflammation-associated depression through NRF2-HO1 signaling

doi: 10.1016/j.ynstr.2026.100804

Figure Lengend Snippet: Deletion of microglial Sirt6 inhibited the NRF2-HO1 signaling and worsened the peroxidation damage. (A) Gene Ontology (GO) analysis was performed on RNA-Seq data from microglia sorted from Sirt6 MCKO and Sirt6 fl/fl control mice. (B) TAC, MDA, SOD, and GSH/GSSG levels at 5 days after LPS injection. n = 4 mice. (C) Gene Set Enrichment Analysis (GSEA) of RNA-Seq data profiled from microglia sorted from Sirt6 MCKO and Sirt6 fl/fl control mice. (D-E) Analysis of NRF2-HO1 and associated signaling proteins in sorted microglia. Protein levels of NRF2, KEAP1, HO-1, NQO1, NLRP3, Cleaved Caspase-3, and Cleaved IL-1β were assessed by Western blot (D) and quantified (E). n = 4 mice. Data are mean ± SEM. Statistical significance between two groups was determined by an unpaired two-tailed Student's t-test.

Article Snippet: Membranes were blocked with 5% non-fat milk in TBST for 1 h and incubated overnight at 4 °C with primary antibodies: SIRT6 (12486, Cell Signaling Technology), NRF2 (ab62352, Abcam), HO-1 (ab68477, Abcam), NQO1 (ab80588, Abcam), KEAP1 (8047, Cell Signaling Technology), NLRP3 (AG-20B-0014, AdipoGen), cleaved caspase-1 (4199, Cell Signaling Technology), cleaved IL-1β (83186, Cell Signaling Technology), and β-actin (A1978, Sigma-Aldrich).

Techniques: RNA Sequencing, Control, Injection, Western Blot, Two Tailed Test

Journal: Neurobiology of Stress

Article Title: Microglial SIRT6 confers protection against neuroinflammation-associated depression through NRF2-HO1 signaling

doi: 10.1016/j.ynstr.2026.100804

Figure Lengend Snippet: Knockout of the Nrf2 in microglial aggravated the depression behavior and facilitated the microglial activation. (A) Experimental timeline depicting the sequential administration of tamoxifen and LPS, followed by behavioral assessment. (B) Western blot analysis confirming the successful knockout of Nrf2 in microglial cells of Nrf2 MCKO mice. n = 3 mice. (C-D) Quantification of the duration of immobility in the (C) TST) and (D) FST. n = 8 mice. (E) Levels of TAC, MDA, SOD, and the GSH/GSSG ratio were measured in Nrf2 MCKO and Nrf2 fl/fl mice 3 days after LPS challenge. (F) Immunostaining of GFAP and IBA1 in the hippocampus area of Nrf2 MCKO and Nrf2 fl/fl mice 3 days after LPS injection. (G-L) Quantification of (G) IBA1 + and (H) GFAP + cell counts, along with microglial morphology parameters: (I) number of branches, (J) average branch length, (K) total branch length, and (L) soma area. n = 4 mice. (M) qPCR analysis of TNF-α, IL-6, and IL-1β mRNA levels in the microglial sorted from the hippocampus brain tissue. n = 4 mice. (N) Western blot analysis was conducted to determine the protein levels of SIRT6 and NRF2 downstream signaling components in primary microglia purified from the mouse hippocampus. n = 4 mice. Data are mean ± SEM. Statistical significance between two groups was determined by an unpaired two-tailed Student's t-test.

Article Snippet: Membranes were blocked with 5% non-fat milk in TBST for 1 h and incubated overnight at 4 °C with primary antibodies: SIRT6 (12486, Cell Signaling Technology), NRF2 (ab62352, Abcam), HO-1 (ab68477, Abcam), NQO1 (ab80588, Abcam), KEAP1 (8047, Cell Signaling Technology), NLRP3 (AG-20B-0014, AdipoGen), cleaved caspase-1 (4199, Cell Signaling Technology), cleaved IL-1β (83186, Cell Signaling Technology), and β-actin (A1978, Sigma-Aldrich).

Techniques: Knock-Out, Activation Assay, Western Blot, Immunostaining, Injection, Purification, Two Tailed Test

Journal: Neurobiology of Stress

Article Title: Microglial SIRT6 confers protection against neuroinflammation-associated depression through NRF2-HO1 signaling

doi: 10.1016/j.ynstr.2026.100804

Figure Lengend Snippet: Sirt6 regulated the microglial activation and the depression behavior through Nrf2-HO1 signaling in vivo (A) Experimental timeline of tamoxifen administration, AAV9-Cx3cr1-Nrf2 injection, LPS challenge, and behavioral tests in Sirt6 MCKO mice. (B) Western blot analysis of NRF2 levels after AAV9-Cx3cr1-Nrf2 injection. n = 3 mice. (C-D) Immobility time in TST (C) and FST (D). n = 8 mice. (E) TAC, MDA, SOD, and GSH/GSSG levels at 5 days after LPS injection. n = 4 mice. (F-L) Immunostaining of GFAP and IBA1 in the hippocampus area at 5 days after LPS injection (F); Quantification of (G) IBA1 + and (H) GFAP + cell counts, along with microglial morphology parameters: (I) number of branches, (J) average branch length, (K) total branch length, and (L) soma area. n = 4 mice. (M) qPCR analysis of TNF-α,IL-6, and IL-1β mRNA levels in the microglial sorted from the hippocampus brain tissue. n = 4 mice. (N) Western blot analysis of NRF2 downstream signaling component protein levels in microglia sorted from the hippocampus. n = 4 mice. Data are mean ± SEM. Statistical significance between two groups was determined by an unpaired two-tailed Student's t-test.

Article Snippet: Membranes were blocked with 5% non-fat milk in TBST for 1 h and incubated overnight at 4 °C with primary antibodies: SIRT6 (12486, Cell Signaling Technology), NRF2 (ab62352, Abcam), HO-1 (ab68477, Abcam), NQO1 (ab80588, Abcam), KEAP1 (8047, Cell Signaling Technology), NLRP3 (AG-20B-0014, AdipoGen), cleaved caspase-1 (4199, Cell Signaling Technology), cleaved IL-1β (83186, Cell Signaling Technology), and β-actin (A1978, Sigma-Aldrich).

Techniques: Activation Assay, In Vivo, Injection, Western Blot, Immunostaining, Two Tailed Test

Journal: Neurobiology of Stress

Article Title: Microglial SIRT6 confers protection against neuroinflammation-associated depression through NRF2-HO1 signaling

doi: 10.1016/j.ynstr.2026.100804

Figure Lengend Snippet: Overexpression of Sirt6 in microglial improved the depression behavior and impeded the microglial activation (A) Experimental timeline depicting tamoxifen administration, followed by tail vein injection of AAV9-Cx3cr1-Sirt6, subsequent LPS challenge, and finally, a series of behavioral tests. (B) Western blot analysis of SIRT6 expression after AAV9-Cx3cr1-Sirt6 injection. n = 3 mice. (C-D) The immobility time was quantified in TST (C) and FST (D). n = 8 mice. (E) TAC, MDA, SOD, and GSH/GSSG levels at 5 days after LPS injection. (F-L) Immunostaining of GFAP and IBA1 in the hippocampus area at 5 days after LPS injection (F); Quantification of (G) IBA1 + and (H) GFAP + cell counts, along with microglial morphology parameters: (I) number of branches, (J) average branch length, (K) total branch length, and (L) soma area. n = 4 mice.(M) qPCR analysis of TNF-α,IL-6, and IL-1β mRNA levels in the microglial sorted from the hippocampus brain tissue. n = 4 mice. Data are mean ± SEM. Statistical significance between two groups was determined by an unpaired two-tailed Student's t-test.

Article Snippet: Membranes were blocked with 5% non-fat milk in TBST for 1 h and incubated overnight at 4 °C with primary antibodies: SIRT6 (12486, Cell Signaling Technology), NRF2 (ab62352, Abcam), HO-1 (ab68477, Abcam), NQO1 (ab80588, Abcam), KEAP1 (8047, Cell Signaling Technology), NLRP3 (AG-20B-0014, AdipoGen), cleaved caspase-1 (4199, Cell Signaling Technology), cleaved IL-1β (83186, Cell Signaling Technology), and β-actin (A1978, Sigma-Aldrich).

Techniques: Over Expression, Activation Assay, Injection, Western Blot, Expressing, Immunostaining, Two Tailed Test

Journal: Neurobiology of Stress

Article Title: Microglial SIRT6 confers protection against neuroinflammation-associated depression through NRF2-HO1 signaling

doi: 10.1016/j.ynstr.2026.100804

Figure Lengend Snippet: UBCS039-mediated SIRT6 activation exerts antidepressant and anti-inflammatory effects (A) Schematic of the experimental timeline for UBCS039 administration, LPS injection, and behavioral testing. (B-C) The immobility time was measured in TST (B) and FST (C). n = 4 mice. (D-G) TAC (D), MDA (E), SOD (F), and GSH/GSSG (G) levels at 5 days after LPS injection. n = 4 mice. (H-I) Immunostaining of GFAP and IBA1 in the hippocampus area at 5 days after LPS injection (H); Quantification of IBA1 + and GFAP + cell counts, along with microglial morphology parameters: number of branches, average branch length, total branch length, and soma area (I). n = 4 mice. (J) qPCR analysis of TNF-α,IL-6, and IL-1β mRNA levels in the microglial sorted from the hippocampus brain tissue. n = 4 mice. Data are mean ± SEM. Statistical significance between two groups was determined by an unpaired two-tailed Student's t-test.

Article Snippet: Membranes were blocked with 5% non-fat milk in TBST for 1 h and incubated overnight at 4 °C with primary antibodies: SIRT6 (12486, Cell Signaling Technology), NRF2 (ab62352, Abcam), HO-1 (ab68477, Abcam), NQO1 (ab80588, Abcam), KEAP1 (8047, Cell Signaling Technology), NLRP3 (AG-20B-0014, AdipoGen), cleaved caspase-1 (4199, Cell Signaling Technology), cleaved IL-1β (83186, Cell Signaling Technology), and β-actin (A1978, Sigma-Aldrich).

Techniques: Activation Assay, Injection, Immunostaining, Two Tailed Test

Journal: Cell Communication and Signaling : CCS

Article Title: Expression, regulation, and multifaceted molecular and biological functions of sirtuin 6 in the porcine endometrium during early pregnancy

doi: 10.1186/s12964-026-02699-1

Figure Lengend Snippet: a-c Localization and expression of SIRT6 at the maternal-conceptus interface during early pregnancy. a Immunohistochemical localization of SIRT6 protein in the endometrium of early pregnant gilts (days 10 to 30) and conceptus trophoblast from days 20 and 30 of pregnancy. Positive immunoreaction was observed in luminal (LE) and glandular (GE) epithelium, stromal (ST) cells of endometrium, as well as in the trophoblast (Tr). Arrows indicate nuclear staining. NC – negative control; DP – day of pregnancy; Scale bars, 20 µm. b Expression of SIRT6 mRNA and protein in the endometrium of early pregnant pigs. c Expression of SIRT6 protein in the nuclear fraction of the endometrium. d Effect of conceptus-derived factors on SIRT6 mRNA and protein expression in endometrial explants. Endometrial slices were treated with estradiol (E2; 0.01 and 0.1 µM), prostaglandin E2 (PGE2; 1 and 10 µM), interferon γ (IFNγ; 10 and 50 ng/mL), interleukin 1β (IL1β; 10 and 50 ng/mL), or conceptus-exposed medium (CEM) for 24 h. Values from qPCR were normalized to the geometric mean of GAPDH and HPRT1 gene expression. Values from densitometric analyses of blots were normalized to total protein content using TGX Stain-Free gel technology. All blots are included in Additional File 3: Fig. S4-S6. b and c Data are expressed as the mean ± SEM ( n = 6 per day in each group). Various letters indicate a significant difference between groups ( p < 0.05). d Data are expressed as the mean ± SEM from 5 experiments and presented as a fold change of control values

Article Snippet: The overnight incubation with polyclonal rabbit anti-SIRT6 antibody (1:50; Cell Signaling Technology, Danvers, MA, USA) was accomplished at 4 °C in a humidified chamber.

Techniques: Expressing, Immunohistochemical staining, Staining, Negative Control, Derivative Assay, Gene Expression, Control

Journal: Cell Communication and Signaling : CCS

Article Title: Expression, regulation, and multifaceted molecular and biological functions of sirtuin 6 in the porcine endometrium during early pregnancy

doi: 10.1186/s12964-026-02699-1

Figure Lengend Snippet: Localization and regulation of SIRT6 expression in luminal epithelial (LE) cells of the porcine endometrium. a Representative images showing SIRT6 protein localization in LE cells. Cells were counterstained with diamidino-2-phenylindole (DAPI) and CytoPainter Phalloidin-iFluor 488 Reagent (iFluor) to visualize nuclei and actin filaments, respectively. NC – negative control; scale bars, 20 µm. Cells were treated with ( b ) estradiol (E2; 0.01 and 0.1 µM), prostaglandin E2 (PGE2; 1 and 10 µM), interferon γ (IFNγ; 10 and 50 ng/mL), interleukin 1β (IL1β; 10 and 50 ng/mL), ( c ) conceptus-exposed medium (CEM), ( d ) E2 (0.1 µM), progesterone (P4; 0.1 µM), or the combination of E2 and P4 for 24 h. Values from qPCR were normalized to the geometric mean of GAPDH and HPRT1 gene expression. Values from densitometric analyses of blots were normalized to the total protein content using TGX Stain-Free gel technology ( b and c ) or to the abundance of beta-actin (ACTB; d ). All blots are included in Additional File 3: Fig. S7. The results are presented as a fold change of the respective control values. Data are expressed as the mean ± SEM from 3–4 ( c ) and 6 ( b and d ) experiments

Article Snippet: The overnight incubation with polyclonal rabbit anti-SIRT6 antibody (1:50; Cell Signaling Technology, Danvers, MA, USA) was accomplished at 4 °C in a humidified chamber.

Techniques: Expressing, Negative Control, Gene Expression, Staining, Control

Journal: Cell Communication and Signaling : CCS

Article Title: Expression, regulation, and multifaceted molecular and biological functions of sirtuin 6 in the porcine endometrium during early pregnancy

doi: 10.1186/s12964-026-02699-1

Figure Lengend Snippet: The transcriptome of porcine endometrial tissue in response to SIRT6 activation. a Volcano plot with differentially expressed genes (DEGs) in endometrial explants treated with SIRT6 activator, UBCS039, compared to untreated control (adjusted p -value < 0.05 and log2 fold change > 0.58). Green dots and red dots represent up-regulated and down-regulated DEGs, respectively. Grey dots represent genes with no significant difference. b Heatmap showing hierarchical clustering analysis results according to DEGs after UBCS039 treatment of endometrial explants; the top 50 DEGs are shown. Each row represents one gene, and each column represents a sample. The color scale indicates the relative gene expression level, expressed as a z-score, which represents the number of standard deviations from the mean expression of a given gene. Green indicates higher values in gene expression, and red indicates lower values compared with the respective control (CTRL). c The qPCR validation of RNA-sequencing (RNA-seq) data using eleven selected genes. Values from qPCR were normalized to the geometric mean of GAPDH and HPRT1 gene expression. Data are expressed as the log2 fold change of mean expression levels between control and UBCS039-treated endometrial explants ( n = 3 per group). TSPAN15 : tetraspanin 15; F11R : F11 receptor; MMP3 : matrix metallopeptidase 3; FLT1 : fms related receptor tyrosine kinase 1; ANGPT1 : angiopoietin 1; ASNS : asparagine synthetase (glutamine-hydrolyzing); SLC3A2 : solute carrier family 3 member 2; TMED3 : transmembrane p24 trafficking protein 3; UBA5 : ubiquitin like modifier activating enzyme 5; SARS1 : Seryl-tRNA synthetase 1; ACAT1 : acetyl-CoA acetyltransferase 1

Article Snippet: The overnight incubation with polyclonal rabbit anti-SIRT6 antibody (1:50; Cell Signaling Technology, Danvers, MA, USA) was accomplished at 4 °C in a humidified chamber.

Techniques: Activation Assay, Control, Gene Expression, Expressing, Biomarker Discovery, RNA Sequencing, Ubiquitin Proteomics

Journal: Cell Communication and Signaling : CCS

Article Title: Expression, regulation, and multifaceted molecular and biological functions of sirtuin 6 in the porcine endometrium during early pregnancy

doi: 10.1186/s12964-026-02699-1

Figure Lengend Snippet: Effect of SIRT6 on prostaglandin (PG) E2 concentration and prostaglandin E synthase ( PTGES ) mRNA expression. a and b Endometrial slices were exposed to medium only (control) or medium containing SIRT6 activator (UBCS039; 75 and 100 µM) or inhibitor (OSS_128167; 100 µM and 125 µM) for 24 h. c Arachidonic acid (ARA; 200 µM) was used as a positive control for PGE2 synthesis. d Luminal epithelial (LE) cells were cultured with UBCS039 (100 µM) for 6 and 24 h. PGE2 levels are presented as a fold change of the respective control values. Values from qPCR were normalized to the geometric mean of GAPDH and HPRT1 gene expression. Data are expressed as the mean ± SEM from 4 ( c , d ) or 5 ( a , b ) experiments. Bars with various letters are significantly different ( p < 0.05). Asterisk indicates the difference compared with the control ( p < 0.05)

Article Snippet: The overnight incubation with polyclonal rabbit anti-SIRT6 antibody (1:50; Cell Signaling Technology, Danvers, MA, USA) was accomplished at 4 °C in a humidified chamber.

Techniques: Concentration Assay, Expressing, Control, Positive Control, Cell Culture, Gene Expression

Journal: Cell Communication and Signaling : CCS

Article Title: Expression, regulation, and multifaceted molecular and biological functions of sirtuin 6 in the porcine endometrium during early pregnancy

doi: 10.1186/s12964-026-02699-1

Figure Lengend Snippet: SIRT6 downregulates the BAX/BCL2 ratio by upregulating the expression of the anti-apoptotic BCL2 gene. Endometrial slices were exposed to medium only (control) or medium containing SIRT6 activator (UBCS039; 75 and 100 µM) or inhibitor (OSS_128167; 125 µM) for 24 h. Values from qPCR were normalized to the geometric mean of GAPDH and HPRT1 gene expression. Data are expressed as the mean ± SEM from 5 experiments. Bars with various letters are significantly different ( p < 0.05)

Article Snippet: The overnight incubation with polyclonal rabbit anti-SIRT6 antibody (1:50; Cell Signaling Technology, Danvers, MA, USA) was accomplished at 4 °C in a humidified chamber.

Techniques: Expressing, Control, Gene Expression

Journal: Cell Communication and Signaling : CCS

Article Title: Expression, regulation, and multifaceted molecular and biological functions of sirtuin 6 in the porcine endometrium during early pregnancy

doi: 10.1186/s12964-026-02699-1

Figure Lengend Snippet: SIRT6 stimulates luminal epithelial (LE) cell proliferation by promoting cell cycle progression. a and b Effect of SIRT6 on LE cell proliferation. Cells were cultured with SIRT6 activator, UBCS039, or inhibitor, OSS_128167 (50 to 125 µM) for 24 and 48 h. Newborn calf serum (NCS; 20%) was used as a positive control. c and d Effect of SIRT6 on cell cycle distribution and the expression of essential regulators of cell cycle progression. LE cells were treated with either control media or UBCS039 (100 µM). c After 24 h, the percentage of cells in G0/G1, S, and G2/M phases was calculated by fluorescence intensity of incorporated FxCycle Violet Stain in a flow cytometry analysis. Representative images of cell cycle fractions are presented. d After 6 and/or 24 h, cell cycle-related protein levels were determined in cell extracts by Western blot. Values from densitometric analyses of blots were normalized to the abundance of beta-actin (ACTB). Western blots used for densitometric quantifications of cyclins are presented. All blots are included in Additional File 3: Fig. S8. Data are expressed as the mean ± SEM ( n = 3–4). Asterisks indicate differences as compared with the respective control cells (* p < 0.05; ** p < 0.01; **** p < 0.0001). Bars with various letters (ab-for control cells; xy-for UBCS039-treated cells) are significantly different among groups

Article Snippet: The overnight incubation with polyclonal rabbit anti-SIRT6 antibody (1:50; Cell Signaling Technology, Danvers, MA, USA) was accomplished at 4 °C in a humidified chamber.

Techniques: Cell Culture, Positive Control, Expressing, Control, Fluorescence, Staining, Flow Cytometry, Western Blot

Journal: Cell Communication and Signaling : CCS

Article Title: Expression, regulation, and multifaceted molecular and biological functions of sirtuin 6 in the porcine endometrium during early pregnancy

doi: 10.1186/s12964-026-02699-1

Figure Lengend Snippet: Activation of SIRT6 disrupts the adhesive properties of endometrial luminal epithelial cells. Cells were treated with either control media or UBCS039 (100 µM). Data are expressed as the mean ± SEM ( n = 3). Asterisk indicates difference as compared with the control cells (* p < 0.05)

Article Snippet: The overnight incubation with polyclonal rabbit anti-SIRT6 antibody (1:50; Cell Signaling Technology, Danvers, MA, USA) was accomplished at 4 °C in a humidified chamber.

Techniques: Activation Assay, Adhesive, Control

Journal: Cell Communication and Signaling : CCS

Article Title: Expression, regulation, and multifaceted molecular and biological functions of sirtuin 6 in the porcine endometrium during early pregnancy

doi: 10.1186/s12964-026-02699-1

Figure Lengend Snippet: Graphical summary of the molecular and biological effects of SIRT6 in the porcine peri-implantation endometrium. Molecules and cellular processes affected by SIRT6 are indicated in green (stimulation) and red (inhibition). A SIRT6 facilitates the transport of serine, glucose, and glutamine via the SLC transporters for metabolism; an intermediate of glycolysis is converted into serine by enzymatic actions of PHGDH, PSAT1, and PSPH, and then into glycine and formate via SHMT2 and MTHFD2; glutamine is converted to α-ketoglutarate (α-KG) by PSAT1 and GPT2, for entry into the TCA cycle. This releases aspartate, which is converted to asparagine via ASNS. SIRT6 may drive the biosynthesis of proline from glutamate by enhancing the activity of ALDH18A1 and PYCR1, and by inhibiting ALDH4A1. B SIRT6 induces the polyamine-spermidine pathway by up-regulating ODC1 and SRM expression and affecting polyamine uptake and transport. C SIRT6 facilitates the import of protein precursors into mitochondria. D SIRT6-affected genes are related to the oxidative phosphorylation (OXPHOS) system and ATP production. E SIRT6 may balance pro- and anti-inflammatory responses; e.g., it inhibits the cytokine-dependent JAK/STAT5/IRF pathway and modulates the levels of anti-inflammatory OSM and pro-inflammatory CSF. SIRT6 regulates the levels of TNF family members and inhibits TRADD, a TNF receptor 1-associated signal transducer. F SIRT6 is essential for intracellular trafficking by facilitating (i) the translocation of proteins across the ER membrane involving the SRP complex, (ii) anterograde and retrograde Golgi trafficking involving COPII and COPI components, and (iii) extracellular vesicle-mediated transport involving RAB proteins. G SIRT6 controls the initiation of translation by inducing the expression of aminoacyl-tRNA-synthetases (aaRS), METTL , and ALKBH1 . H SIRT6 affects extracellular matrix remodeling and impairs adhesion of luminal epithelial cells. I Anti-apoptotic action of SIRT6. J SIRT6 accelerates cell cycle progression through the G2/M phase and promotes cell proliferation. K SIRT6 induces prostaglandin (PG) E2 metabolism. The full names of depicted molecules are listed in Additional file 1: Table S3

Article Snippet: The overnight incubation with polyclonal rabbit anti-SIRT6 antibody (1:50; Cell Signaling Technology, Danvers, MA, USA) was accomplished at 4 °C in a humidified chamber.

Techniques: Inhibition, Activity Assay, Expressing, Phospho-proteomics, Translocation Assay, Membrane

Journal: Nature Communications

Article Title: Sirt6 deficiency in mast cells promotes adipose fibroinflammation in obesity through galectin-3 signaling

doi: 10.1038/s41467-025-66040-z

Figure Lengend Snippet: a Representative immunofluorescence staining of mast cells in omental adipose tissue from individuals with a BMI < 25 (lean), 25 ≤ BMI < 30 (overweight), and BMI ≥ 30 (obesity). Both total and degranulated mast cells were counted ( n = 13 for lean, n = 16 for overweight, n = 25 for obese). b Adipose-derived mast cells (FcεRI + CD117 + ) from the SVF were isolated from human omental adipose tissues, and Sirt6 protein levels were compared among individuals with a BMI < 25, 25 ≤ BMI < 30, and BMI ≥ 30 ( n = 25 for lean, n = 14 for overweight, n = 6 for obese). c A scatter plot was created to compare mast cell Sirt6 expression with body mass index (BMI, n = 45), plasma leptin ( n = 44), and plasma adiponectin ( n = 44) levels. d , e C57BL/6 mice were fed either a normal chow diet (NCD) or a high-fat diet (HFD) for 3 or 16 weeks. Epididymal adipose tissues were then harvested for western blot analysis of Sirt6 in mast cells ( d ) or for histological analysis with toluidine blue staining to identify mast cells ( e ). Both total and degranulated mast cells were counted ( n = 14 for NCD, n = 15 for HFD). Values are presented as mea n ± SD. One-way ANOVA followed by Sidak’s multiple comparisons analysis (a, b) and unpaired two-tailed t test between two groups ( e ) were conducted for statistical analyses. Pearson correlation coefficients were calculated between continuous variables in two-sided distributions ( c ). Source data are provided as a Source Data file.

Article Snippet: Chromatin was immunoprecipitated overnight at 4 °C with antibodies against Sirt6 (#12486, 1:250 dilution), NF-κB (#8242, 1:250 dilution), nonspecific IgG (#2729, 1:250 dilution, all from Cell Signaling Technology), and Ac-H3K9 (#H9286, 1:250 dilution, Sigma-Aldrich). qPCR was performed on ChIP DNA to assess the occupancy of Sirt6 or Ac-H3K9 at the Lgals3 and Lgals9 promoters.

Techniques: Immunofluorescence, Staining, Derivative Assay, Isolation, Expressing, Clinical Proteomics, Western Blot, Two Tailed Test

Journal: Nature Communications

Article Title: Sirt6 deficiency in mast cells promotes adipose fibroinflammation in obesity through galectin-3 signaling

doi: 10.1038/s41467-025-66040-z

Figure Lengend Snippet: a Cma1-Cre (Cre), WT ( Sirt6 fl/fl , fl/fl) and KO ( Sirt6 -/- , -/-) mice were fed a HFD for 16 weeks from the age of 6 weeks and metabolically assessed. b , c Body weight changes (b, n = 8 per group) and food intake (c, n = 8 per group). d , e Glucose concentrations during an intraperitoneal glucose tolerance ( d , n = 8 per group) and an insulin tolerance test ( e , n = 8 per group) were measured, and areas under the curve were compared. f Fat and liver tissue weights at the end of study ( n = 8 per group). g Adipose tissue sections were H&E stained, and adipocyte size in H&E sections was quantified ( n = 52 for Cre, n = 55 for WT, n = 55 for KO). Values are presented as mean ± SD. Values on the line in panels ( b , d , e ) represent statistical comparisons with the Sirt6 fl/fl group. Two-way ANOVA followed by Bonferroni’s post hoc analysis ( b – e ) and one-way ANOVA followed by Sidak’s multiple comparisons analysis was conducted for statistical analyses ( f , g ). Source data are provided as a Source Data file. Panel ( a ) was created in BioRender. H, J. (2025) https://BioRender.com/lh1p3yb .

Article Snippet: Chromatin was immunoprecipitated overnight at 4 °C with antibodies against Sirt6 (#12486, 1:250 dilution), NF-κB (#8242, 1:250 dilution), nonspecific IgG (#2729, 1:250 dilution, all from Cell Signaling Technology), and Ac-H3K9 (#H9286, 1:250 dilution, Sigma-Aldrich). qPCR was performed on ChIP DNA to assess the occupancy of Sirt6 or Ac-H3K9 at the Lgals3 and Lgals9 promoters.

Techniques: Metabolic Labelling, Staining

Journal: Nature Communications

Article Title: Sirt6 deficiency in mast cells promotes adipose fibroinflammation in obesity through galectin-3 signaling

doi: 10.1038/s41467-025-66040-z

Figure Lengend Snippet: a . Schematic illustration of the transfer of WT or Sirt6-deficient adipose-derived mast cells (ADMCs, 2 ×10 6 cells) into mast cell-deficient Kit W-sh/W-sh mice. b - e . Body weights changes over time (b, n = 7 per group), wet weight of epididymal (EAT), inguinal (IAT), brown adipose tissue (BAT), and liver (c, n = 7 per group), glucose concentrations during an intraperitoneal glucose tolerance (d, n = 7 per group) and an insulin tolerance test (e, n = 7 per group) were measured. Areas under the curve during glucose and insulin tolerance tests were compared. f . EAT sections were stained with H&E or immunostained with antibodies against tryptase. Adipocyte size in H&E-stained sections ( n = 37 per group) and total and degranulated mast cell numbers in tryptase immunostained sections ( n = 4 per group) were determined. Values are presented as mean ± SD. Red and blue values on the line in panels b, d, and e indicate statistical comparisons with the Sirt6 fl/fl mast cell group and PBS group, respectively. One-way ANOVA followed by Sidak’s multiple comparisons analysis (c, f) and two-way ANOVA followed by Bonferroni’s post hoc analysis (b, d, e) were conducted for statistical analyses Source data are provided as a Source Data file. Panel (a) was created in BioRender. H, J. (2025) https://BioRender.com/49artbl .

Article Snippet: Chromatin was immunoprecipitated overnight at 4 °C with antibodies against Sirt6 (#12486, 1:250 dilution), NF-κB (#8242, 1:250 dilution), nonspecific IgG (#2729, 1:250 dilution, all from Cell Signaling Technology), and Ac-H3K9 (#H9286, 1:250 dilution, Sigma-Aldrich). qPCR was performed on ChIP DNA to assess the occupancy of Sirt6 or Ac-H3K9 at the Lgals3 and Lgals9 promoters.

Techniques: Derivative Assay, Staining

Journal: Nature Communications

Article Title: Sirt6 deficiency in mast cells promotes adipose fibroinflammation in obesity through galectin-3 signaling

doi: 10.1038/s41467-025-66040-z

Figure Lengend Snippet: a , b mRNA levels (a, n = 6 per group) in EAT tissues and protein levels (b, n = 6 per group) in plasma for galectin-1, galectin-3 and galectin-9 in WT and KO mice. c Heatmap displaying the ligand-receptor signaling network of the galectin pathway between immune cell types from WT and KO mice. Each cell in the heatmap represents the predicted communication probability from a sender to a receiver cell type. The bar plot above the heatmap quantifies the total outgoing signaling strength of each cell type, while the bar plot to the right reflects the total incoming signaling strength. TCP, total communication probability. d Bone marrow-derived mast cells (BMMCs) were transfected with control siRNA (siCtrl) or Sirt6 siRNA (siSirt6). Successful Sirt6 silencing was confirmed by qPCR and western blotting ( n = 6 per group). e – i Lgals3 and Lgals9 mRNA levels (e, n = 6 per group), RELA (p65 subunit of NF-κB) binding to the promoters of Lgals3 and Lgals9 ( f , n = 5 per group), Sirt6 binding to the promoters of Lgals3 and Lgasl9 ( g , n = 5 per group), enrichment of Ac-H3K9 on the Lgals3 and Lgals9 promoters ( h , n = 4 per group), and acetylation of H3K9 ( i , n = 3 per group) were compared between control and Sirt6 silenced BMMCs. j , k . Plasma levels of galectin-3 and galectin-9 in human subjects were measured by ELISA ( n = 24 for lean, n = 14 for overweight, n = 6 for obese). A scatter plot was created to examine the correlation between mast cell Sirt6 protein levels and plasma galectin-3 and galectin-9 levels. Values are presented as mean ± SD. Unpaired two-tailed t test between the two groups ( a , b , d , e , i ), one-way ANOVA followed by Sidak’s multiple comparisons analysis ( j , k ) and two-way ANOVA followed by Bonferroni’s post hoc analysis were conducted for statistical analyses ( f – h ). Pearson correlation coefficients were calculated between continuous variables in two-sided distributions ( j , k). Source data are provided as a Source Data file.

Article Snippet: Chromatin was immunoprecipitated overnight at 4 °C with antibodies against Sirt6 (#12486, 1:250 dilution), NF-κB (#8242, 1:250 dilution), nonspecific IgG (#2729, 1:250 dilution, all from Cell Signaling Technology), and Ac-H3K9 (#H9286, 1:250 dilution, Sigma-Aldrich). qPCR was performed on ChIP DNA to assess the occupancy of Sirt6 or Ac-H3K9 at the Lgals3 and Lgals9 promoters.

Techniques: Clinical Proteomics, Derivative Assay, Transfection, Control, Western Blot, Binding Assay, Enzyme-linked Immunosorbent Assay, Two Tailed Test

Journal: Nature Communications

Article Title: Sirt6 deficiency in mast cells promotes adipose fibroinflammation in obesity through galectin-3 signaling

doi: 10.1038/s41467-025-66040-z

Figure Lengend Snippet: a Schematic illustration of the adoptive transfer of WT or Sirt6-deficient adipose-derived mast cells (ADMCs), infected with lentivirus carrying shRNA targeting either control shRNA (Lv-eGFP-shCtrl, shCtrl) or galectin-3 (Lv-eGFP-shLgals3, shLgals3), into Kit W-sh/W-sh mice. b – d Body weights during 16 weeks of HFD ( b , n = 6 per group), glucose concentrations during an intraperitoneal glucose tolerance ( c , n = 6 per group) and an insulin tolerance test ( d , n = 6 per group). Areas under the curve during glucose and insulin tolerance tests were compared. Red and pink values on the line in panels ( b , c , and d ) indicate statistical comparisons with the Sirt6 fl/fl MC+shCtrl and Sirt6 -/- MC+shCtrl group, respectively. e EAT sections were stained with H&E or Sirius Red, or immunostained with anti-F4/80 antibody. Representative images and quantification results of adipocyte size in H&E-stained sections ( n = 37 per group) and macrophage numbers in F4/80-immunostained sections ( n = 7 per group) are shown. f Hydroxyproline content in EAT of mice as a measure of fibrosis ( f , n = 6 per group). g EAT sections were immunostained for tryptase, and total and degranulated mast cell counts ( n = 4 per group) were determined ( g ). Values are presented as mean ± SD. Two-way ANOVA followed by Bonferroni’s post hoc analysis ( b – d ) and one-way ANOVA followed by Sidak’s multiple comparisons analysis ( e – g ) were conducted for statistical analyses. Source data are provided as a Source Data file. Panel ( a ) was created in BioRender. H, J. (2025) https://BioRender.com/swlgc9x .

Article Snippet: Chromatin was immunoprecipitated overnight at 4 °C with antibodies against Sirt6 (#12486, 1:250 dilution), NF-κB (#8242, 1:250 dilution), nonspecific IgG (#2729, 1:250 dilution, all from Cell Signaling Technology), and Ac-H3K9 (#H9286, 1:250 dilution, Sigma-Aldrich). qPCR was performed on ChIP DNA to assess the occupancy of Sirt6 or Ac-H3K9 at the Lgals3 and Lgals9 promoters.

Techniques: Adoptive Transfer Assay, Derivative Assay, Infection, shRNA, Control, Staining