primary antibody against sirt6  (Cell Signaling Technology Inc)

Journal: Cell Death & Disease

Article Title: Impaired SUMOylation of FoxA1 promotes nonalcoholic fatty liver disease through down-regulation of Sirt6

doi: 10.1038/s41419-024-07054-1

Figure Lengend Snippet: A RT-qPCR analysis of Sirt6, Acox1, Lpl, Pparα, Cpt1a, and Cyp4a14 levels in the liver tissues of mice. B The protein levels of Sirt6, FoxO1, Ac-FoxO1, Pparα, Acox1, Lpl, Cpt1a, and Cyp4a14 in livers were determined by Western blotting. C Primary hepatocytes were isolated from FoxA1-LKO mice and infected with Ad-LacZ or Ad-FoxA1 before treatment with palmitic acid (PAL) (0.4 mM). Lipid droplet formation was evaluated by BODIPY staining (scale bar = 100 µm). D , E RT-qPCR and Western blotting analyzed Sirt6, FoxO1, Ac-FoxO1, Pparα, Acox1, Lpl, Cpt1a and Cyp4a14 expression in the primary hepatocytes. F , G Primary hepatocytes were isolated from FoxA1 flox/flox mice and infected with Ad-shLacZ or Ad-shFoxA1, followed by treatment with PAL. Sirt6, FoxO1, Ac-FoxO1, Pparα, Acox1, Lpl, Cpt1a, and Cyp4a14 expression levels were measured by RT-qPCR and Western blotting. H , I Primary hepatocytes from FoxA1 flox/flox mice were infected with Ad-FoxA1, Ad-shSirt6, or a combination of them, and then treated with PAL. Sirt6, FoxO1, Ac-FoxO1, Pparα, Acox1, Lpl, Cpt1a, and Cyp4a14 expression levels were detected by RT-qPCR and Western blotting. Data was repeated at least 3 times. * p < 0.05, ** p < 0.01, and *** p < 0.001. For A-G, Student’s t test was performed. For H-I, one-way ANOVA followed by Tukey’s multiple comparison test was performed.

Article Snippet: The blots were blocked by skim milk and incubated using the primary antibodies against Sirt6 (A18468, 1:500, ABclonal), Cpt1a (ab234111, 1:1000, Abcam), Acox1 (ab184032, 1:1000, Abcam), Lpl (ab91606, 1:1000, Abcam), Pparα (bs-3614R, 1:500, Bioss), Cyp4a14 (ab3573, 1:1000, Abcam), FoxA1 (A15278, 1:500, ABclonal), Sae1 (ab185949, 1:1000, Abcam), Sae2 (ab185955, 1:1000, Abcam), Ubc9 (ab33044, 1:1000, Abcam), Pias1/2 (ab77231, 1:1000, Abcam), Pias3 (ab105178, 1:1000, Abcam), Pias4 (ab137500, 1:1000, Abcam), FoxO1 (ab70382, 1:2000, Abcam), Ac-FoxO1 (PA5-104560, 1:1000, Thermo Fisher), Ac-K (ab190479, 1:1000, Abcam), Cpt2 (ab181114, 1:1000, Abcam), β-actin (bs-0061R, 1:5000, Bioss) at 4 °C overnight.

Techniques: Quantitative RT-PCR, Western Blot, Isolation, Infection, Staining, Expressing, Comparison

Journal: Cell Death & Disease

Article Title: Impaired SUMOylation of FoxA1 promotes nonalcoholic fatty liver disease through down-regulation of Sirt6

doi: 10.1038/s41419-024-07054-1

Figure Lengend Snippet: Primary hepatocytes extracted from FoxA1-LKO mice were transduced with Ad-LacZ or Ad-Sirt6, followed by PAL treatment. A Primary hepatocytes were received BODIPY staining to observe lipid droplets (scale bar = 100 µm). After feeding with HFD for 2 weeks, Ad-LacZ were injected into FoxA1 flox/flox mice, and Ad-LacZ or Ad-Sirt6 were injected into FoxA1-LKO mice. 14 days after injection, the liver tissues were collected. B , C Sirt6, FoxO1, Ac-FoxO1, Pparα, Acox1 and Lpl levels were detected by RT-qPCR and Western blotting, respectively. D , E Liver cholesterol and triglycerides levels were determined. F Liver steatosis was evaluated by HE staining (scale bar = 50 µm). G Lipid accumulation in livers was analyzed by Oil-Red O staining (scale bar = 50 µm). N = 6, * p < 0.05, ** p < 0.01, and *** p < 0.001. One-way ANOVA followed by Tukey’s multiple comparison test was performed.

Article Snippet: The blots were blocked by skim milk and incubated using the primary antibodies against Sirt6 (A18468, 1:500, ABclonal), Cpt1a (ab234111, 1:1000, Abcam), Acox1 (ab184032, 1:1000, Abcam), Lpl (ab91606, 1:1000, Abcam), Pparα (bs-3614R, 1:500, Bioss), Cyp4a14 (ab3573, 1:1000, Abcam), FoxA1 (A15278, 1:500, ABclonal), Sae1 (ab185949, 1:1000, Abcam), Sae2 (ab185955, 1:1000, Abcam), Ubc9 (ab33044, 1:1000, Abcam), Pias1/2 (ab77231, 1:1000, Abcam), Pias3 (ab105178, 1:1000, Abcam), Pias4 (ab137500, 1:1000, Abcam), FoxO1 (ab70382, 1:2000, Abcam), Ac-FoxO1 (PA5-104560, 1:1000, Thermo Fisher), Ac-K (ab190479, 1:1000, Abcam), Cpt2 (ab181114, 1:1000, Abcam), β-actin (bs-0061R, 1:5000, Bioss) at 4 °C overnight.

Techniques: Transduction, Staining, Injection, Quantitative RT-PCR, Western Blot, Comparison

Journal: Cell Death & Disease

Article Title: Impaired SUMOylation of FoxA1 promotes nonalcoholic fatty liver disease through down-regulation of Sirt6

doi: 10.1038/s41419-024-07054-1

Figure Lengend Snippet: The primary hepatocytes were treated with 0.4 mM palmitic acid (PAL) for 48 h. A The sites of SUMOylated FoxA1 was predicted by sumoplot. B Western blotting analysis of Sae1, Sae2, Ubc9, Pias1/2, Pias3, Pias4 protein levels in primary hepatocytes. C Ni 2+ -NTA pull-down assay was performed to validate the interaction between FoxA1 and Sumo1, Sumo2, or Sumo3 in HEK293T cells. D Ni 2+ -NTA pull-down assay was performed to validate the interaction between FoxA1 and Sumo2 in AML-12 cells. E The direct binding of Sumo2 to FoxA1 protein in primary hepatocytes was evaluated by Co-IP. Mice were subjected to regular chow or HFD feeding for 10 weeks. F FoxA1 SUMOylation level in livers was detected by Co-IP assay. Lipid droplet formation in primary hepatocytes was observed by Oil red O staining ( G ) and BODIPY staining ( H ) (scale bar = 100 µm). I Western blotting analysis of protein levels of Sirt6 and Pparα in primary hepatocytes. Data was repeated at least 3 times. ** p < 0.01. Student’s t test was performed.

Article Snippet: The blots were blocked by skim milk and incubated using the primary antibodies against Sirt6 (A18468, 1:500, ABclonal), Cpt1a (ab234111, 1:1000, Abcam), Acox1 (ab184032, 1:1000, Abcam), Lpl (ab91606, 1:1000, Abcam), Pparα (bs-3614R, 1:500, Bioss), Cyp4a14 (ab3573, 1:1000, Abcam), FoxA1 (A15278, 1:500, ABclonal), Sae1 (ab185949, 1:1000, Abcam), Sae2 (ab185955, 1:1000, Abcam), Ubc9 (ab33044, 1:1000, Abcam), Pias1/2 (ab77231, 1:1000, Abcam), Pias3 (ab105178, 1:1000, Abcam), Pias4 (ab137500, 1:1000, Abcam), FoxO1 (ab70382, 1:2000, Abcam), Ac-FoxO1 (PA5-104560, 1:1000, Thermo Fisher), Ac-K (ab190479, 1:1000, Abcam), Cpt2 (ab181114, 1:1000, Abcam), β-actin (bs-0061R, 1:5000, Bioss) at 4 °C overnight.

Techniques: Western Blot, Pull Down Assay, Binding Assay, Co-Immunoprecipitation Assay, Staining

Journal: Cell Death & Disease

Article Title: Impaired SUMOylation of FoxA1 promotes nonalcoholic fatty liver disease through down-regulation of Sirt6

doi: 10.1038/s41419-024-07054-1

Figure Lengend Snippet: Primary hepatocytes and AML-12 cells were infected with Ad-FoxA1, and then stimulated with 0.4 mM PAL for 48 h. A FoxA1 and Sirt6 protein levels were measured by Western blotting. B JASPAR database predicted the binding sites of FoxA1 to Sirt6 promoter. C ChIP assay validated the direct interaction between FoxA1 and Sirt6 promoter. D The transcription activity of Sirt6 was detected by dual-luciferase reporter assay. PAL-treated primary hepatocytes were further administrated with SUMOylation inhibitor (GA, 10 nM) or activator (N106, 10 μM). E FoxA1 SUMOylation was assessed by the SUMOylation Assay Ultra Kit. F Ni-NTA pull-down assay was performed to determine the interaction between FoxA1 and Sumo2. G Western blotting analysis of Sirt6, Pparα, and Cpt2 protein levels and RT-qPCR analysis of Pparα, Cpt2 and Cpt1a in primary hepatocytes. H Lipid droplet formation was evaluated by BODIPY staining (scale bar = 100 µm). Data was repeated at least 3 times. * p < 0.05, ** p < 0.01, and *** p < 0.001. For ( C ), Student’s t test was performed. For ( A , D , E , G , H ), one-way ANOVA followed by Tukey’s multiple comparison test was performed.

Article Snippet: The blots were blocked by skim milk and incubated using the primary antibodies against Sirt6 (A18468, 1:500, ABclonal), Cpt1a (ab234111, 1:1000, Abcam), Acox1 (ab184032, 1:1000, Abcam), Lpl (ab91606, 1:1000, Abcam), Pparα (bs-3614R, 1:500, Bioss), Cyp4a14 (ab3573, 1:1000, Abcam), FoxA1 (A15278, 1:500, ABclonal), Sae1 (ab185949, 1:1000, Abcam), Sae2 (ab185955, 1:1000, Abcam), Ubc9 (ab33044, 1:1000, Abcam), Pias1/2 (ab77231, 1:1000, Abcam), Pias3 (ab105178, 1:1000, Abcam), Pias4 (ab137500, 1:1000, Abcam), FoxO1 (ab70382, 1:2000, Abcam), Ac-FoxO1 (PA5-104560, 1:1000, Thermo Fisher), Ac-K (ab190479, 1:1000, Abcam), Cpt2 (ab181114, 1:1000, Abcam), β-actin (bs-0061R, 1:5000, Bioss) at 4 °C overnight.

Techniques: Infection, Western Blot, Binding Assay, Activity Assay, Luciferase, Reporter Assay, Pull Down Assay, Quantitative RT-PCR, Staining, Comparison

Journal: Cell Death & Disease

Article Title: Impaired SUMOylation of FoxA1 promotes nonalcoholic fatty liver disease through down-regulation of Sirt6

doi: 10.1038/s41419-024-07054-1

Figure Lengend Snippet: A Conserved FoxA1 SUMOylation sites at lysine residue K6. B HEK293T cells were transfected with WT-FoxA1 or FoxA1 mutants (K6R, K266R, K388R) in combination with His-Sumo2, Flag-Ubc9. Ni-NTA pull-down assay evaluated the interaction between FoxA1 and Sumo2. AML-12 cells were transfected with WT-FoxA1 or FoxA1 K6R with or without treatment with PAL. C , D RT-qPCR and Western blotting detected Sirt6 and Pparα expression. E Dual-luciferase reporter assay determined the transcription activity of Sirt6. F SUMOylation Assay Ultra Kit measured FoxA1 SUMOylation level. G The transcription activity of Sirt6 in hepatocytes with indicated treatments was measured by dual-luciferase reporter assay. Data was repeated at least 3 times. * p < 0.05, ** p < 0.01, and *** p < 0.001. For ( C – G ), One-way ANOVA followed by Tukey’s multiple comparison test was performed.

Article Snippet: The blots were blocked by skim milk and incubated using the primary antibodies against Sirt6 (A18468, 1:500, ABclonal), Cpt1a (ab234111, 1:1000, Abcam), Acox1 (ab184032, 1:1000, Abcam), Lpl (ab91606, 1:1000, Abcam), Pparα (bs-3614R, 1:500, Bioss), Cyp4a14 (ab3573, 1:1000, Abcam), FoxA1 (A15278, 1:500, ABclonal), Sae1 (ab185949, 1:1000, Abcam), Sae2 (ab185955, 1:1000, Abcam), Ubc9 (ab33044, 1:1000, Abcam), Pias1/2 (ab77231, 1:1000, Abcam), Pias3 (ab105178, 1:1000, Abcam), Pias4 (ab137500, 1:1000, Abcam), FoxO1 (ab70382, 1:2000, Abcam), Ac-FoxO1 (PA5-104560, 1:1000, Thermo Fisher), Ac-K (ab190479, 1:1000, Abcam), Cpt2 (ab181114, 1:1000, Abcam), β-actin (bs-0061R, 1:5000, Bioss) at 4 °C overnight.

Techniques: Residue, Transfection, Pull Down Assay, Quantitative RT-PCR, Western Blot, Expressing, Luciferase, Reporter Assay, Activity Assay, Comparison

Journal: Cell Death & Disease

Article Title: Impaired SUMOylation of FoxA1 promotes nonalcoholic fatty liver disease through down-regulation of Sirt6

doi: 10.1038/s41419-024-07054-1

Figure Lengend Snippet: A The primary hepatocytes were transfected with Ad-shMdm2, Ad-shMdm4, Ad-shUbox5, and FoxA1 protein levels were determined by Western blotting. B , C Co-IP confirmed the exogenous and endogenous interplay between Mdm2 and FoxA1 proteins. D The primary hepatocytes were treated with PAL together with or without MG132 (100 nM). Western blotting analysis of FoxA1 protein level. E The primary hepatocytes were infected with Ad-Mdm2 or Ad-NC, followed by treatment with PAL or/ and MG132. FoxA1 protein level was detected by Western blotting. F The primary hepatocytes were infected with Ad-shMdm2 or Ad-shNC, and then exposed to CHX for 1, 2, 4 h. Western blotting analysis of FoxA1 protein level. G AML-12 cells were transfected with WT-FoxA1 or FoxA1 K6R, and then exposed to CHX for 1, 2, 4 h. Western blotting analysis of FoxA1 protein level. H , I The ubiquitylation level of FoxA1 in primary hepatocytes and AML-12 cells was evaluated by Co-IP. J The protein levels of Mdm2, FoxA1, Sirt6 and Pparα in primary hepatocytes were determined by Western blotting. K Lipid droplet formation was observed by BODIPY staining (scale bar = 100 µm). Data was repeated at least 3 times. * p < 0.05, ** p < 0.01, and *** p < 0.001. For ( A , F , G ), Student’s t test was performed. For ( D , E , J – K ), One-way ANOVA followed by Tukey’s multiple comparison test was performed.

Article Snippet: The blots were blocked by skim milk and incubated using the primary antibodies against Sirt6 (A18468, 1:500, ABclonal), Cpt1a (ab234111, 1:1000, Abcam), Acox1 (ab184032, 1:1000, Abcam), Lpl (ab91606, 1:1000, Abcam), Pparα (bs-3614R, 1:500, Bioss), Cyp4a14 (ab3573, 1:1000, Abcam), FoxA1 (A15278, 1:500, ABclonal), Sae1 (ab185949, 1:1000, Abcam), Sae2 (ab185955, 1:1000, Abcam), Ubc9 (ab33044, 1:1000, Abcam), Pias1/2 (ab77231, 1:1000, Abcam), Pias3 (ab105178, 1:1000, Abcam), Pias4 (ab137500, 1:1000, Abcam), FoxO1 (ab70382, 1:2000, Abcam), Ac-FoxO1 (PA5-104560, 1:1000, Thermo Fisher), Ac-K (ab190479, 1:1000, Abcam), Cpt2 (ab181114, 1:1000, Abcam), β-actin (bs-0061R, 1:5000, Bioss) at 4 °C overnight.

Techniques: Transfection, Western Blot, Co-Immunoprecipitation Assay, Infection, Staining, Comparison

Journal: Cell Death & Disease

Article Title: Impaired SUMOylation of FoxA1 promotes nonalcoholic fatty liver disease through down-regulation of Sirt6

doi: 10.1038/s41419-024-07054-1

Figure Lengend Snippet: The mice were treated with GA or N106 during HFD or normal diet feeding for 12 weeks. A , B Liver weight, white adipose tissue (WAT) weight, brown adipose tissue (BAT), inguinal white adipose tissue (iWAT), and epigonadal white adipose tissue (eWAT) were measured. C Blood glucose levels of mice was detected against time after glucose or insulin injection. D – F Serum ALT, liver cholesterol and triglycerides levels were detected. G HE staining determined the pathological changes in livers (scale bar = 50 µm). H FoxA1, Sirt6, and Pparα expression in liver tissues was evaluated by immunohistochemical staining (scale bar = 50 µm). Primary hepatocytes were isolated from mice in different groups. I RT-qPCR analysis of Cd36, Fasn, Dgat, Adipoq, Fabp4, Cebpa, Pparg, Atgl, Cpt1a and Cpt2 mRNA levels in livers from mice treated with GA or N106 during HFD or normal diet feeding for 12 weeks. J BODIPY staining detected lipid droplet formation (scale bar = 100 µm). K FoxA1 SUMOylation level was measured by SUMOylation Assay Ultra Kit. ( L ) Western blotting analysis of FoxA1, Sirt6, and Pparα protein abundance. N = 6, * p < 0.05, ** p < 0.01, and *** p < 0.001. For ( C ), ANOVA for repeated measurement was performed; For ( A , B , D – F , I – L ), One-way ANOVA followed by Tukey’s multiple comparison test was performed.

Article Snippet: The blots were blocked by skim milk and incubated using the primary antibodies against Sirt6 (A18468, 1:500, ABclonal), Cpt1a (ab234111, 1:1000, Abcam), Acox1 (ab184032, 1:1000, Abcam), Lpl (ab91606, 1:1000, Abcam), Pparα (bs-3614R, 1:500, Bioss), Cyp4a14 (ab3573, 1:1000, Abcam), FoxA1 (A15278, 1:500, ABclonal), Sae1 (ab185949, 1:1000, Abcam), Sae2 (ab185955, 1:1000, Abcam), Ubc9 (ab33044, 1:1000, Abcam), Pias1/2 (ab77231, 1:1000, Abcam), Pias3 (ab105178, 1:1000, Abcam), Pias4 (ab137500, 1:1000, Abcam), FoxO1 (ab70382, 1:2000, Abcam), Ac-FoxO1 (PA5-104560, 1:1000, Thermo Fisher), Ac-K (ab190479, 1:1000, Abcam), Cpt2 (ab181114, 1:1000, Abcam), β-actin (bs-0061R, 1:5000, Bioss) at 4 °C overnight.

Techniques: Injection, Staining, Expressing, Immunohistochemical staining, Isolation, Quantitative RT-PCR, Western Blot, Quantitative Proteomics, Comparison

Journal: Immunity, Inflammation and Disease

Article Title: X‐Box binding protein 1 downregulates SIRT6 to promote injury in pancreatic ductal epithelial cells

doi: 10.1002/iid3.1301

Figure Lengend Snippet: X‐Box binding protein 1 (XBP1) negatively regulates SIRT6. (A) Site of XBP1 binding to the SIRT6 promoter according to the JASPAR website. (B) Reverse transcription‐quantitative polymerase chain reaction (RT‐qPCR) reveals XBP1 level upon caerulein treatment. (C) Western blot analysis reveals XBP1, GRP78, and CHOP protein levels upon caerulein treatment. (D) RT‐qPCR and (E) western blot analysis reveal XBP1 level after transfection. (F) RT‐qPCR and (G) western blot analysis reveal SIRT6 level in response to XBP1 overexpression or knockdown. (H) Luciferase assay was used to determine SIRT6 promoter activity. (I) Binding of XBP1 to the SIRT6 promoter was determined using chromatin immunoprecipitation (ChIP). (J) The level of CHOP in human pancreatic duct epithelial (HPDE) cells was determined using immunofluorescence assay, magnification 200×. ** p < .01, *** p < .001 versus control or oe‐NC or lgG; ### p < .001 versus sh‐NC.

Article Snippet: Next, membranes were blocked with 5% skimmed milk, followed by the incubation with primary antibodies against SIRT6, XBP1, ER stress‐related, apoptosis‐related proteins and HRP‐conjugated secondary antibody (Proteintech group or Abcam).

Techniques: Binding Assay, Reverse Transcription, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Western Blot, Transfection, Over Expression, Knockdown, Luciferase, Activity Assay, Chromatin Immunoprecipitation, Immunofluorescence, Control

Journal: Immunity, Inflammation and Disease

Article Title: X‐Box binding protein 1 downregulates SIRT6 to promote injury in pancreatic ductal epithelial cells

doi: 10.1002/iid3.1301

Figure Lengend Snippet: X‐Box binding protein 1 (XBP1) curbs the impacts of SIRT6 on the inflammation and oxidative stress. Cells were subjected to simultaneous transfection with SIRT6 and XBP1 overexpression plasmids. (A) the levels of inflammatory factors in the cell supernatant were measured upon caerulein treatment. (B) The dichlorodihydrofluorescein diacetate (DCFH‐DA) probe was used to quantify reactive oxygen species (ROS) levels in human pancreatic duct epithelial (HPDE) cells, magnification 200×. (C) Oxidative stress was assessed by measuring malondialdehyde (MDA) level, superoxide dismutase (SOD), and catalase (CAT) activities. *** p < .001 versus control; ### p < .001 versus caerulein; @ p < .05, p @ @ < .01, @@@ p < .001 versus caerulein + oe‐SIRT6 + oe‐NC.

Article Snippet: Next, membranes were blocked with 5% skimmed milk, followed by the incubation with primary antibodies against SIRT6, XBP1, ER stress‐related, apoptosis‐related proteins and HRP‐conjugated secondary antibody (Proteintech group or Abcam).

Techniques: Binding Assay, Transfection, Over Expression, Control

Journal: Immunity, Inflammation and Disease

Article Title: X‐Box binding protein 1 downregulates SIRT6 to promote injury in pancreatic ductal epithelial cells

doi: 10.1002/iid3.1301

Figure Lengend Snippet: X‐Box binding protein 1 (XBP1) curbs the impacts of SIRT6 on the apoptosis. Cells were subjected to simultaneous transfection with SIRT6 and XBP1 overexpression plasmids. (A) Flow cytometry was used to detect cell apoptosis. (B) Caspase 3 activity was also used to assess apoptosis. (C) Western blot analysis reveals the protein levels of Bcl‐2 and Bax. *** p < .001 versus control; ### p < .001 versus caerulein; @ p < .05, p @ @ < .01, p @ @ @ < .001 versus caerulein + oe‐SIRT6 + oe‐NC.

Article Snippet: Next, membranes were blocked with 5% skimmed milk, followed by the incubation with primary antibodies against SIRT6, XBP1, ER stress‐related, apoptosis‐related proteins and HRP‐conjugated secondary antibody (Proteintech group or Abcam).

Techniques: Binding Assay, Transfection, Over Expression, Flow Cytometry, Activity Assay, Western Blot, Control

Journal: Anais brasileiros de dermatologia

Article Title: Expression analysis and biological regulation of silencing regulatory protein 6 (SIRT6) in cutaneous squamous cell carcinoma.

doi: 10.1016/j.abd.2023.08.010

Figure Lengend Snippet: Figure 1 Expression of SIRT6 in NT (Normal skin Tissue), AK (Actinic Keratosis), BD (Bowen Disease), and CSCC (Cutaneous Squamous Cell Carcinoma) tissues. (A) Representative images (magnified by ×100 × 200) of SIRT6 immunohistochemical staining in NT, AK, BD,

Article Snippet: The authors hypothesize that SIRT6 plays a pivotal ole in the pathogenesis and progression of CSCC, prompting s to evaluate its expression patterns, functional mechaisms, and potential clinical implications. aterials and methods aterials eagents he following reagents were used: A431 human cutaneous quamous cell carcinoma cell line (Obtained from Zhongqiao inzhou, Shanghai) and SCL-1 cell line (Enzyme Research, hanghai); The normal immortalized human keratinocyte ine HaCaT (Shanghai Fuheng Biotechnology Co., Ltd.);skim ilk, chloroform, and isopropanol (purchased from Shanghai henggong Co., Ltd.); primary antibodies against SIRT6 and -tubulin, secondary antibodies against mouse immunogloblin IgG (purchased from Abcam, USA); mouse anti-human -cadherin monoclonal antibody and mouse anti-human imentin monoclonal antibody (Proteintech, Wuhan); fetal ovine serum and high-glucose DMEM medium (purchased rom Gibco, USA); BCA analysis kit, crystal violet, and Lipoectamine 2000 (purchased from Biyuntian Biotechnology o., Ltd.); Polyvinylidene Fluoride (PVDF) membrane (purhased from Millipore, USA); Trizol reagent, RNA prep pure issue Kit, PrimeScript RT synthesis kit, and SYBR Green qRTCR Master Mix kit (purchased from Thermo, USA); CCK-8 ssay kit (Proteintech, Wuhan); matrix gel and 8- m tran- a g H s PRESS .

Techniques: Expressing, Immunohistochemical staining, Staining

Journal: Anais brasileiros de dermatologia

Article Title: Expression analysis and biological regulation of silencing regulatory protein 6 (SIRT6) in cutaneous squamous cell carcinoma.

doi: 10.1016/j.abd.2023.08.010

Figure Lengend Snippet: Figure 2 SIRT6 knockdown inhibited CSCC cell proliferation. (A) Western blot analysis of SIRT6 protein expression in A431, SCL-1, and

Article Snippet: The authors hypothesize that SIRT6 plays a pivotal ole in the pathogenesis and progression of CSCC, prompting s to evaluate its expression patterns, functional mechaisms, and potential clinical implications. aterials and methods aterials eagents he following reagents were used: A431 human cutaneous quamous cell carcinoma cell line (Obtained from Zhongqiao inzhou, Shanghai) and SCL-1 cell line (Enzyme Research, hanghai); The normal immortalized human keratinocyte ine HaCaT (Shanghai Fuheng Biotechnology Co., Ltd.);skim ilk, chloroform, and isopropanol (purchased from Shanghai henggong Co., Ltd.); primary antibodies against SIRT6 and -tubulin, secondary antibodies against mouse immunogloblin IgG (purchased from Abcam, USA); mouse anti-human -cadherin monoclonal antibody and mouse anti-human imentin monoclonal antibody (Proteintech, Wuhan); fetal ovine serum and high-glucose DMEM medium (purchased rom Gibco, USA); BCA analysis kit, crystal violet, and Lipoectamine 2000 (purchased from Biyuntian Biotechnology o., Ltd.); Polyvinylidene Fluoride (PVDF) membrane (purhased from Millipore, USA); Trizol reagent, RNA prep pure issue Kit, PrimeScript RT synthesis kit, and SYBR Green qRTCR Master Mix kit (purchased from Thermo, USA); CCK-8 ssay kit (Proteintech, Wuhan); matrix gel and 8- m tran- a g H s PRESS .

Techniques: Knockdown, Western Blot, Expressing

Journal: Anais brasileiros de dermatologia

Article Title: Expression analysis and biological regulation of silencing regulatory protein 6 (SIRT6) in cutaneous squamous cell carcinoma.

doi: 10.1016/j.abd.2023.08.010

Figure Lengend Snippet: Figure 3 Inhibition of CSCC Cell Migration and Invasion by SIRT6 knockdown. (A) Transwell assay results and statistical analysis representing

Article Snippet: The authors hypothesize that SIRT6 plays a pivotal ole in the pathogenesis and progression of CSCC, prompting s to evaluate its expression patterns, functional mechaisms, and potential clinical implications. aterials and methods aterials eagents he following reagents were used: A431 human cutaneous quamous cell carcinoma cell line (Obtained from Zhongqiao inzhou, Shanghai) and SCL-1 cell line (Enzyme Research, hanghai); The normal immortalized human keratinocyte ine HaCaT (Shanghai Fuheng Biotechnology Co., Ltd.);skim ilk, chloroform, and isopropanol (purchased from Shanghai henggong Co., Ltd.); primary antibodies against SIRT6 and -tubulin, secondary antibodies against mouse immunogloblin IgG (purchased from Abcam, USA); mouse anti-human -cadherin monoclonal antibody and mouse anti-human imentin monoclonal antibody (Proteintech, Wuhan); fetal ovine serum and high-glucose DMEM medium (purchased rom Gibco, USA); BCA analysis kit, crystal violet, and Lipoectamine 2000 (purchased from Biyuntian Biotechnology o., Ltd.); Polyvinylidene Fluoride (PVDF) membrane (purhased from Millipore, USA); Trizol reagent, RNA prep pure issue Kit, PrimeScript RT synthesis kit, and SYBR Green qRTCR Master Mix kit (purchased from Thermo, USA); CCK-8 ssay kit (Proteintech, Wuhan); matrix gel and 8- m tran- a g H s PRESS .

Techniques: Inhibition, Migration, Knockdown, Transwell Assay

Journal: Anais brasileiros de dermatologia

Article Title: Expression analysis and biological regulation of silencing regulatory protein 6 (SIRT6) in cutaneous squamous cell carcinoma.

doi: 10.1016/j.abd.2023.08.010

Figure Lengend Snippet: Figure 4 SIRT6 knockdown can increase apoptosis and cell cycle arrest in G0/G1 phase of CSCC cells. (A---B) Flow cytomatics analysis showed

Article Snippet: The authors hypothesize that SIRT6 plays a pivotal ole in the pathogenesis and progression of CSCC, prompting s to evaluate its expression patterns, functional mechaisms, and potential clinical implications. aterials and methods aterials eagents he following reagents were used: A431 human cutaneous quamous cell carcinoma cell line (Obtained from Zhongqiao inzhou, Shanghai) and SCL-1 cell line (Enzyme Research, hanghai); The normal immortalized human keratinocyte ine HaCaT (Shanghai Fuheng Biotechnology Co., Ltd.);skim ilk, chloroform, and isopropanol (purchased from Shanghai henggong Co., Ltd.); primary antibodies against SIRT6 and -tubulin, secondary antibodies against mouse immunogloblin IgG (purchased from Abcam, USA); mouse anti-human -cadherin monoclonal antibody and mouse anti-human imentin monoclonal antibody (Proteintech, Wuhan); fetal ovine serum and high-glucose DMEM medium (purchased rom Gibco, USA); BCA analysis kit, crystal violet, and Lipoectamine 2000 (purchased from Biyuntian Biotechnology o., Ltd.); Polyvinylidene Fluoride (PVDF) membrane (purhased from Millipore, USA); Trizol reagent, RNA prep pure issue Kit, PrimeScript RT synthesis kit, and SYBR Green qRTCR Master Mix kit (purchased from Thermo, USA); CCK-8 ssay kit (Proteintech, Wuhan); matrix gel and 8- m tran- a g H s PRESS .

Techniques: Knockdown