polyclonal rabbit anti sirt6 antibody  (Cell Signaling Technology Inc)

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: medRxiv

Article Title: Quercetin activates the SIRT6–Nrf2 axis during oxidative stress, modulating DNA repair and ageing-associated markers

doi: 10.1101/2025.10.31.25338366

Figure Lengend Snippet: Fold change in A) SIRT1; B) SIRT6; C) PARP1; D) OGG1; E) RAD51; F) XRCC1 comparing placebo (gray bars) vs quercetin (green bars). Orange asterisk indicates significant within-subject difference.

Article Snippet: Cells were fixed with 4 % paraformaldehyde in PBS (Sigma-Aldrich) for 10 min, washed three times in PBS (Sigma-Aldrich), and permeabilised with 0.5 % Triton X-100/PBS (Sigma-Aldrich) for 10 min. Non-specific binding was blocked with 2.5 % normal horse serum (NHS; Vector Laboratories, Burlingame, CA, USA) for 1 h. Slides were then incubated with rabbit polyclonal anti-SIRT6 antibody (1:500; Proteintech, 28677-1-AP, Manchester, UK) in 1 % NHS/PBS for 1 h at room temperature, washed three times in 0.05 % Tween-20/1 % NHS/PBS, and incubated with goat anti-rabbit Alexa Fluor 568 antibody (1:500; Invitrogen, Thermo Fisher Scientific) for 1 h. Quercetin autofluorescence was visualised directly in the green channel (excitation 488 nm), eliminating the need for secondary staining.

Techniques:

Journal: medRxiv

Article Title: Quercetin activates the SIRT6–Nrf2 axis during oxidative stress, modulating DNA repair and ageing-associated markers

doi: 10.1101/2025.10.31.25338366

Figure Lengend Snippet: A) SIRT6 colocalisation with native, unstained quercetin (%ROI colocalised) comparing placebo (gray bars) vs quercetin (green bars); B) Confocal microscopy of colocalisation. Black asterisk indicates significant between-group difference. Orange asterisk indicates significant within-subject difference.

Article Snippet: Cells were fixed with 4 % paraformaldehyde in PBS (Sigma-Aldrich) for 10 min, washed three times in PBS (Sigma-Aldrich), and permeabilised with 0.5 % Triton X-100/PBS (Sigma-Aldrich) for 10 min. Non-specific binding was blocked with 2.5 % normal horse serum (NHS; Vector Laboratories, Burlingame, CA, USA) for 1 h. Slides were then incubated with rabbit polyclonal anti-SIRT6 antibody (1:500; Proteintech, 28677-1-AP, Manchester, UK) in 1 % NHS/PBS for 1 h at room temperature, washed three times in 0.05 % Tween-20/1 % NHS/PBS, and incubated with goat anti-rabbit Alexa Fluor 568 antibody (1:500; Invitrogen, Thermo Fisher Scientific) for 1 h. Quercetin autofluorescence was visualised directly in the green channel (excitation 488 nm), eliminating the need for secondary staining.

Techniques: Confocal Microscopy

Journal: Practical laboratory medicine

Article Title: Impact of hemolysis on the levels of proteins associated with aging and age-related neurodegenerative diseases in a multicentric clinical research.

doi: 10.1016/j.plabm.2025.e00455

Figure Lengend Snippet: Fig. 3. Difference in protein concentrations in Non-Hemolyzed versus Hemolyzed samples: (a) Comparable concentration can be observed for SIRT1 protein. (b) A significant increase of 1.11ng/ul was observed for SIRT2 protein in Hemolyzed blood plasma compared to non-hemolyzed samples (p < 0.0001). (c) Increase in SIRT6 protein was not found to be significantly affected due to hemolysis (p = 0.0564). (d) A significant decline of 1.29ng/ul was observed for FOXO3A protein in hemolyzed blood plasma compared to non-hemolyzed samples (p = 0.0272).

Article Snippet: Primary antibodies used were mouse anti-human SIRT1 monoclonal IgG (Sc-74504,Santa Cruz Biotechnology Inc. USA), goat anti-human SIRT2 polyclonal IgG (Sc-31912, Santa Cruz Biotechnology Inc. USA), rabbit anti-human SIRT6 polyclonal IgG (Sc517196,Santa Cruz Biotechnology Inc. USA), mouse anti-human FOXO3A IgG (Sc-48348,Santa Cruz Biotechnology Inc. USA), goat anti-human t-Tau IgG (Sc-32274, Santa Cruz Biotechnology Inc. USA) and rabbit anti-human p-Tau181 IgG (12885S (D9F4G), Cell Signaling Technology Inc. MA, USA) were immobilized in different flow cells of CM5 sensor chip via amine coupling kit (Wipro GE Healthcare, Sweden).

Techniques: Concentration Assay, Clinical Proteomics

Journal: Practical Laboratory Medicine

Article Title: Impact of hemolysis on the levels of proteins associated with aging and age-related neurodegenerative diseases in a multicentric clinical research

doi: 10.1016/j.plabm.2025.e00455

Figure Lengend Snippet: Quantitative comparison of proteins in Hemolyzed and Non Hemolyzed plasma samples by SPR.

Article Snippet: Primary antibodies used were mouse anti-human SIRT1 monoclonal IgG (Sc-74504,Santa Cruz Biotechnology Inc. USA), goat anti-human SIRT2 polyclonal IgG (Sc-31912, Santa Cruz Biotechnology Inc. USA), rabbit anti-human SIRT6 polyclonal IgG (Sc-517196,Santa Cruz Biotechnology Inc. USA), mouse anti-human FOXO3A IgG (Sc-48348,Santa Cruz Biotechnology Inc. USA), goat anti-human t-Tau IgG (Sc-32274, Santa Cruz Biotechnology Inc. USA) and rabbit anti-human p-Tau181 IgG (12885S (D9F4G), Cell Signaling Technology Inc. MA, USA) were immobilized in different flow cells of CM5 sensor chip via amine coupling kit (Wipro GE Healthcare, Sweden).

Techniques: Comparison, Clinical Proteomics, Concentration Assay

Journal: Practical Laboratory Medicine

Article Title: Impact of hemolysis on the levels of proteins associated with aging and age-related neurodegenerative diseases in a multicentric clinical research

doi: 10.1016/j.plabm.2025.e00455

Figure Lengend Snippet: Difference in protein concentrations in Non-Hemolyzed versus Hemolyzed samples: (a) Comparable concentration can be observed for SIRT1 protein. (b) A significant increase of 1.11ng/ul was observed for SIRT2 protein in Hemolyzed blood plasma compared to non-hemolyzed samples (p < 0.0001). (c) Increase in SIRT6 protein was not found to be significantly affected due to hemolysis (p = 0.0564). (d) A significant decline of 1.29ng/ul was observed for FOXO3A protein in hemolyzed blood plasma compared to non-hemolyzed samples (p = 0.0272).

Article Snippet: Primary antibodies used were mouse anti-human SIRT1 monoclonal IgG (Sc-74504,Santa Cruz Biotechnology Inc. USA), goat anti-human SIRT2 polyclonal IgG (Sc-31912, Santa Cruz Biotechnology Inc. USA), rabbit anti-human SIRT6 polyclonal IgG (Sc-517196,Santa Cruz Biotechnology Inc. USA), mouse anti-human FOXO3A IgG (Sc-48348,Santa Cruz Biotechnology Inc. USA), goat anti-human t-Tau IgG (Sc-32274, Santa Cruz Biotechnology Inc. USA) and rabbit anti-human p-Tau181 IgG (12885S (D9F4G), Cell Signaling Technology Inc. MA, USA) were immobilized in different flow cells of CM5 sensor chip via amine coupling kit (Wipro GE Healthcare, Sweden).

Techniques: Concentration Assay, Clinical Proteomics

Journal: Renal Failure

Article Title: Sirt6 overexpression relieves ferroptosis and delays the progression of diabetic nephropathy via Nrf2/GPX4 pathway

doi: 10.1080/0886022x.2024.2377785

Figure Lengend Snippet: Figure 3. Expression of Sirt6 in renal tissues of db/m and db/db 16w group. (A–B) The proteins level of Sirt6, ACSL4, Nrf2, HO1, SOD2 and GPX4 were analyzed by Western blot in renal tissue of db/m and db/db group and shown using a histogram. (C)The mRNA levels of Sirt6, Nrf2, GPX4 were determined by real-time qPCR in renal tissues of db/m group and db/db group and shown using a histogram. (D) Immunohistochemical staining for Sitr6 in renal tissue of db/m group and db/db group. (E) The expression of Sirt6 in podocytes from db/db mice and db/m mice by immunofluorescence, WT-1 was used as podocyte marker. db/m: normal control group; db/db: 16w diabetic group; *p < 0.05, **p < 0.01 db/db vs db/m.

Article Snippet: Immunofluorescence assay Podocytes grown on cover slides were fixed with 4% formaldehyde for 30 min at room temperature, permeabilized 10 min with PbS containing 0.1% triton X-100, blocked for 10% fetal calf serum for 30 min at 37 °C. the samples were incubated with rabbit anti-Sirt6 polyclonal antibody (1:100, 13572-1-aP, Proteintech, Wuhan, China), rabbit anti-Nrf2 (1:50, 16396-1-aP, Proteintech), mouse anti-GPX4 (1:100, 67763-1-Ig, Proteintech), rabbit anti-Sod2 (1:50, 24127-1-aP, Proteintech), rabbit anti-Ho1 (1:200, 10701-1-aP, Proteintech) overnight at 4 °C, and then incubated with goat anti-rabbit or goat anti-mouse IgG H&L secondary antibody at 37 °C for 2h.

Techniques: Expressing, Western Blot, Immunohistochemical staining, Staining, Immunofluorescence, Marker, Control

Journal: Renal Failure

Article Title: Sirt6 overexpression relieves ferroptosis and delays the progression of diabetic nephropathy via Nrf2/GPX4 pathway

doi: 10.1080/0886022x.2024.2377785

Figure Lengend Snippet: Figure 4. Expression of Sirt6, Nrf2, GPX4, HO1 and SOD2 in HPC cells after stimulation with 30 mM D-glucose at various time points. (A–H) The protein levels were analyzed by Western blot and shown using a histogram. (I–K) The mRNA levels of Sirt6, Nrf2, GPX4 were determined by quantitative real-time PCR in HPC cells after stimulation with 30 mM D-glucose at various time points and shown using a histogram. (L) Immunofluorescence for Sirt6, Nrf2, GPX4, HO1, SOD2 in HPC cells in NG and HG (×400). *p < 0.05, **p < 0.01.

Article Snippet: Immunofluorescence assay Podocytes grown on cover slides were fixed with 4% formaldehyde for 30 min at room temperature, permeabilized 10 min with PbS containing 0.1% triton X-100, blocked for 10% fetal calf serum for 30 min at 37 °C. the samples were incubated with rabbit anti-Sirt6 polyclonal antibody (1:100, 13572-1-aP, Proteintech, Wuhan, China), rabbit anti-Nrf2 (1:50, 16396-1-aP, Proteintech), mouse anti-GPX4 (1:100, 67763-1-Ig, Proteintech), rabbit anti-Sod2 (1:50, 24127-1-aP, Proteintech), rabbit anti-Ho1 (1:200, 10701-1-aP, Proteintech) overnight at 4 °C, and then incubated with goat anti-rabbit or goat anti-mouse IgG H&L secondary antibody at 37 °C for 2h.

Techniques: Expressing, Western Blot, Real-time Polymerase Chain Reaction, Immunofluorescence

Journal: Renal Failure

Article Title: Sirt6 overexpression relieves ferroptosis and delays the progression of diabetic nephropathy via Nrf2/GPX4 pathway

doi: 10.1080/0886022x.2024.2377785

Figure Lengend Snippet: Figure 6. Sirt6 overexpression attenuated HG induced oxidative stress, mitochondrial dysfunction, ferroptosis and podocyte injury. (A–F) Podocytes were transfected with pcDNA3.1, pcDNA3.1 Sirt6, Scrambled siRNA, Sirt6 siRNA and then stimulated with HG (30 mM) for 48h. Western blots and quantitative analysis showing the relative protein level of Sirt6 after pcDNA3.1 Sirt6 and Sirt6 siRNA transfection of podocytes in different groups. (G–I) Membrane potential of mitochondrial in HPC cells were detected by JC-1 probe. (J) Morphology of mitochondria in HPC cells were detected by mito tracker red probe. *p < 0.05, **p < 0.01 vs NG.

Article Snippet: Immunofluorescence assay Podocytes grown on cover slides were fixed with 4% formaldehyde for 30 min at room temperature, permeabilized 10 min with PbS containing 0.1% triton X-100, blocked for 10% fetal calf serum for 30 min at 37 °C. the samples were incubated with rabbit anti-Sirt6 polyclonal antibody (1:100, 13572-1-aP, Proteintech, Wuhan, China), rabbit anti-Nrf2 (1:50, 16396-1-aP, Proteintech), mouse anti-GPX4 (1:100, 67763-1-Ig, Proteintech), rabbit anti-Sod2 (1:50, 24127-1-aP, Proteintech), rabbit anti-Ho1 (1:200, 10701-1-aP, Proteintech) overnight at 4 °C, and then incubated with goat anti-rabbit or goat anti-mouse IgG H&L secondary antibody at 37 °C for 2h.

Techniques: Over Expression, Transfection, Western Blot, Membrane

Journal: Renal Failure

Article Title: Sirt6 overexpression relieves ferroptosis and delays the progression of diabetic nephropathy via Nrf2/GPX4 pathway

doi: 10.1080/0886022x.2024.2377785

Figure Lengend Snippet: Figure 7. Regulation of Sirt6 expression and analysis of Nrf2, GPX4, Nephrin in HPC cells. (A)The protein levels of Sirt6 in different transfection conditions detected by Western blot. 1.NG; 2. Lipofectamin 3000 2.5 μl + pcDNA3.1 Sirt6 2 μg + P3000 4 μl; 3. Lipofectamin 3000 3.5 μl + pcDNA3.1 Sirt6 2 μg + P3000 4 μl; 4. Lipofectamin 3000 3.5 μl + pcDNA3.1 Sirt6 4 μg + P3000 4 μl. *p < 0.05, **p < 0.01 vs NG. (B–I) The expression of Nrf2, GPX4 and Nephrin in HPC cells detected by Western blot and PCR. (J) Double immunofluorescence staining of Sirt6 (green) and GPX4 (red) in HPC cells (×400). NG: normal glucose; HG: high glucose; HG + Sirt6 + ML385: HG + pcDNA3.1 Sirt6 + ML385 (5 μM); HG + Sirt6: HG + pcDNA3.1 Sirt6. *p < 0.05, **p < 0.01.

Article Snippet: Immunofluorescence assay Podocytes grown on cover slides were fixed with 4% formaldehyde for 30 min at room temperature, permeabilized 10 min with PbS containing 0.1% triton X-100, blocked for 10% fetal calf serum for 30 min at 37 °C. the samples were incubated with rabbit anti-Sirt6 polyclonal antibody (1:100, 13572-1-aP, Proteintech, Wuhan, China), rabbit anti-Nrf2 (1:50, 16396-1-aP, Proteintech), mouse anti-GPX4 (1:100, 67763-1-Ig, Proteintech), rabbit anti-Sod2 (1:50, 24127-1-aP, Proteintech), rabbit anti-Ho1 (1:200, 10701-1-aP, Proteintech) overnight at 4 °C, and then incubated with goat anti-rabbit or goat anti-mouse IgG H&L secondary antibody at 37 °C for 2h.

Techniques: Expressing, Transfection, Western Blot, Double Immunofluorescence Staining

Journal: iScience

Article Title: YZL-51N functions as a selective inhibitor of SIRT7 by NAD + competition to impede DNA damage repair

doi: 10.1016/j.isci.2024.110014

Figure Lengend Snippet:

Article Snippet: Rabbit polyclonal anti-SIRT6 , Cell Signaling Technology , Cat#12486S; RRID: AB_2636969.

Techniques: Virus, Recombinant, Membrane, Giemsa Stain, Activity Assay, Bicinchoninic Acid Protein Assay, Western Blot, Microscopy, Sequencing, Mutagenesis, Software