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SIRT3 is transcriptionally down-regulated by KRAS G12D . A, differential gene expression of nicotinate and nicotinamide metabolism pathways. B, SIRT3 mRNA levels of HPNE cells with KRAS/Off or KRAS/On for 3 h to 96 h. The data are presented as mean ± SD ( n = 3). p values were determined by Student’s t test. ∗∗∗, p < 0.001; ∗∗∗∗, p < 0.0001. C, SIRT3 and KRAS protein levels in HPNE cells with KRAS/Off or KRAS/On for 3 h to 96 h. Cell extracts were analyzed by Western blotting using tubulin as the loading control. Note that the images of KRAS and tubulin bands were derived from the same source images shown in , C and D , as they were from the same experiment. D, effect of <t>SCH772984</t> on expression of ERK1/2, phosphorylated ERK1/2, and SIRT3 protein levels in KRAS/On HPNE cells. Cells were treated with SCH772984 at indicated concentrations for 24 h. Cell extracts were analyzed by Western blotting using vinculin as the loading control. HPNE KRAS/Off cell extracts were used for comparison. E, effect of SCH772984 on expression of ERK1/2, phosphorylated ERK1/2, and SIRT3 protein levels in KRAS/On HEK293 cells. Cells were treated with SCH772984 at indicated concentrations for 24 h. Cell extracts were analyzed by Western blotting using vinculin as the loading control. HEK293 KRAS/Off cell extracts were used for comparison. F, schematic illustration of the promoter activity assay to identify potential KRAS regulated region in the SIRT3 promoter. G, activity of truncated SIRT3 promoters in HPNE KRAS/Off and HPNE KRAS/On (12 h) cells. The data are presented as mean ± SD ( n = 3). p values were determined by Student’s t test. ∗, p < 0.05;∗∗, p < 0.01. HPNE: human pancreatic normal epithelial cell.
Erk Phosphorylation Inhibitor Sch772984, supplied by MedChemExpress, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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phosphorylated erk af1018  (R&D Systems)
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SIRT3 is transcriptionally down-regulated by KRAS G12D . A, differential gene expression of nicotinate and nicotinamide metabolism pathways. B, SIRT3 mRNA levels of HPNE cells with KRAS/Off or KRAS/On for 3 h to 96 h. The data are presented as mean ± SD ( n = 3). p values were determined by Student’s t test. ∗∗∗, p < 0.001; ∗∗∗∗, p < 0.0001. C, SIRT3 and KRAS protein levels in HPNE cells with KRAS/Off or KRAS/On for 3 h to 96 h. Cell extracts were analyzed by Western blotting using tubulin as the loading control. Note that the images of KRAS and tubulin bands were derived from the same source images shown in , C and D , as they were from the same experiment. D, effect of <t>SCH772984</t> on expression of ERK1/2, phosphorylated ERK1/2, and SIRT3 protein levels in KRAS/On HPNE cells. Cells were treated with SCH772984 at indicated concentrations for 24 h. Cell extracts were analyzed by Western blotting using vinculin as the loading control. HPNE KRAS/Off cell extracts were used for comparison. E, effect of SCH772984 on expression of ERK1/2, phosphorylated ERK1/2, and SIRT3 protein levels in KRAS/On HEK293 cells. Cells were treated with SCH772984 at indicated concentrations for 24 h. Cell extracts were analyzed by Western blotting using vinculin as the loading control. HEK293 KRAS/Off cell extracts were used for comparison. F, schematic illustration of the promoter activity assay to identify potential KRAS regulated region in the SIRT3 promoter. G, activity of truncated SIRT3 promoters in HPNE KRAS/Off and HPNE KRAS/On (12 h) cells. The data are presented as mean ± SD ( n = 3). p values were determined by Student’s t test. ∗, p < 0.05;∗∗, p < 0.01. HPNE: human pancreatic normal epithelial cell.
Phosphorylated Erk Af1018, supplied by R&D Systems, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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phosphorylated erk  (Proteintech)
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SIRT3 is transcriptionally down-regulated by KRAS G12D . A, differential gene expression of nicotinate and nicotinamide metabolism pathways. B, SIRT3 mRNA levels of HPNE cells with KRAS/Off or KRAS/On for 3 h to 96 h. The data are presented as mean ± SD ( n = 3). p values were determined by Student’s t test. ∗∗∗, p < 0.001; ∗∗∗∗, p < 0.0001. C, SIRT3 and KRAS protein levels in HPNE cells with KRAS/Off or KRAS/On for 3 h to 96 h. Cell extracts were analyzed by Western blotting using tubulin as the loading control. Note that the images of KRAS and tubulin bands were derived from the same source images shown in , C and D , as they were from the same experiment. D, effect of <t>SCH772984</t> on expression of ERK1/2, phosphorylated ERK1/2, and SIRT3 protein levels in KRAS/On HPNE cells. Cells were treated with SCH772984 at indicated concentrations for 24 h. Cell extracts were analyzed by Western blotting using vinculin as the loading control. HPNE KRAS/Off cell extracts were used for comparison. E, effect of SCH772984 on expression of ERK1/2, phosphorylated ERK1/2, and SIRT3 protein levels in KRAS/On HEK293 cells. Cells were treated with SCH772984 at indicated concentrations for 24 h. Cell extracts were analyzed by Western blotting using vinculin as the loading control. HEK293 KRAS/Off cell extracts were used for comparison. F, schematic illustration of the promoter activity assay to identify potential KRAS regulated region in the SIRT3 promoter. G, activity of truncated SIRT3 promoters in HPNE KRAS/Off and HPNE KRAS/On (12 h) cells. The data are presented as mean ± SD ( n = 3). p values were determined by Student’s t test. ∗, p < 0.05;∗∗, p < 0.01. HPNE: human pancreatic normal epithelial cell.
Phosphorylated Erk, supplied by Proteintech, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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SIRT3 is transcriptionally down-regulated by KRAS G12D . A, differential gene expression of nicotinate and nicotinamide metabolism pathways. B, SIRT3 mRNA levels of HPNE cells with KRAS/Off or KRAS/On for 3 h to 96 h. The data are presented as mean ± SD ( n = 3). p values were determined by Student’s t test. ∗∗∗, p < 0.001; ∗∗∗∗, p < 0.0001. C, SIRT3 and KRAS protein levels in HPNE cells with KRAS/Off or KRAS/On for 3 h to 96 h. Cell extracts were analyzed by Western blotting using tubulin as the loading control. Note that the images of KRAS and tubulin bands were derived from the same source images shown in , C and D , as they were from the same experiment. D, effect of <t>SCH772984</t> on expression of ERK1/2, phosphorylated ERK1/2, and SIRT3 protein levels in KRAS/On HPNE cells. Cells were treated with SCH772984 at indicated concentrations for 24 h. Cell extracts were analyzed by Western blotting using vinculin as the loading control. HPNE KRAS/Off cell extracts were used for comparison. E, effect of SCH772984 on expression of ERK1/2, phosphorylated ERK1/2, and SIRT3 protein levels in KRAS/On HEK293 cells. Cells were treated with SCH772984 at indicated concentrations for 24 h. Cell extracts were analyzed by Western blotting using vinculin as the loading control. HEK293 KRAS/Off cell extracts were used for comparison. F, schematic illustration of the promoter activity assay to identify potential KRAS regulated region in the SIRT3 promoter. G, activity of truncated SIRT3 promoters in HPNE KRAS/Off and HPNE KRAS/On (12 h) cells. The data are presented as mean ± SD ( n = 3). p values were determined by Student’s t test. ∗, p < 0.05;∗∗, p < 0.01. HPNE: human pancreatic normal epithelial cell.
Phosphorylated Erk (Thr202/Tyr204) Antibody, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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SIRT3 is transcriptionally down-regulated by KRAS G12D . A, differential gene expression of nicotinate and nicotinamide metabolism pathways. B, SIRT3 mRNA levels of HPNE cells with KRAS/Off or KRAS/On for 3 h to 96 h. The data are presented as mean ± SD ( n = 3). p values were determined by Student’s t test. ∗∗∗, p < 0.001; ∗∗∗∗, p < 0.0001. C, SIRT3 and KRAS protein levels in HPNE cells with KRAS/Off or KRAS/On for 3 h to 96 h. Cell extracts were analyzed by Western blotting using tubulin as the loading control. Note that the images of KRAS and tubulin bands were derived from the same source images shown in , C and D , as they were from the same experiment. D, effect of <t>SCH772984</t> on expression of ERK1/2, phosphorylated ERK1/2, and SIRT3 protein levels in KRAS/On HPNE cells. Cells were treated with SCH772984 at indicated concentrations for 24 h. Cell extracts were analyzed by Western blotting using vinculin as the loading control. HPNE KRAS/Off cell extracts were used for comparison. E, effect of SCH772984 on expression of ERK1/2, phosphorylated ERK1/2, and SIRT3 protein levels in KRAS/On HEK293 cells. Cells were treated with SCH772984 at indicated concentrations for 24 h. Cell extracts were analyzed by Western blotting using vinculin as the loading control. HEK293 KRAS/Off cell extracts were used for comparison. F, schematic illustration of the promoter activity assay to identify potential KRAS regulated region in the SIRT3 promoter. G, activity of truncated SIRT3 promoters in HPNE KRAS/Off and HPNE KRAS/On (12 h) cells. The data are presented as mean ± SD ( n = 3). p values were determined by Student’s t test. ∗, p < 0.05;∗∗, p < 0.01. HPNE: human pancreatic normal epithelial cell.
Phosphorylated P Erk, supplied by Proteintech, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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anti phosphorylated p erk  (Proteintech)
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SIRT3 is transcriptionally down-regulated by KRAS G12D . A, differential gene expression of nicotinate and nicotinamide metabolism pathways. B, SIRT3 mRNA levels of HPNE cells with KRAS/Off or KRAS/On for 3 h to 96 h. The data are presented as mean ± SD ( n = 3). p values were determined by Student’s t test. ∗∗∗, p < 0.001; ∗∗∗∗, p < 0.0001. C, SIRT3 and KRAS protein levels in HPNE cells with KRAS/Off or KRAS/On for 3 h to 96 h. Cell extracts were analyzed by Western blotting using tubulin as the loading control. Note that the images of KRAS and tubulin bands were derived from the same source images shown in , C and D , as they were from the same experiment. D, effect of <t>SCH772984</t> on expression of ERK1/2, phosphorylated ERK1/2, and SIRT3 protein levels in KRAS/On HPNE cells. Cells were treated with SCH772984 at indicated concentrations for 24 h. Cell extracts were analyzed by Western blotting using vinculin as the loading control. HPNE KRAS/Off cell extracts were used for comparison. E, effect of SCH772984 on expression of ERK1/2, phosphorylated ERK1/2, and SIRT3 protein levels in KRAS/On HEK293 cells. Cells were treated with SCH772984 at indicated concentrations for 24 h. Cell extracts were analyzed by Western blotting using vinculin as the loading control. HEK293 KRAS/Off cell extracts were used for comparison. F, schematic illustration of the promoter activity assay to identify potential KRAS regulated region in the SIRT3 promoter. G, activity of truncated SIRT3 promoters in HPNE KRAS/Off and HPNE KRAS/On (12 h) cells. The data are presented as mean ± SD ( n = 3). p values were determined by Student’s t test. ∗, p < 0.05;∗∗, p < 0.01. HPNE: human pancreatic normal epithelial cell.
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rabbit anti mouse phosphorylated erk  (Cell Signaling Technology Inc)
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SIRT3 is transcriptionally down-regulated by KRAS G12D . A, differential gene expression of nicotinate and nicotinamide metabolism pathways. B, SIRT3 mRNA levels of HPNE cells with KRAS/Off or KRAS/On for 3 h to 96 h. The data are presented as mean ± SD ( n = 3). p values were determined by Student’s t test. ∗∗∗, p < 0.001; ∗∗∗∗, p < 0.0001. C, SIRT3 and KRAS protein levels in HPNE cells with KRAS/Off or KRAS/On for 3 h to 96 h. Cell extracts were analyzed by Western blotting using tubulin as the loading control. Note that the images of KRAS and tubulin bands were derived from the same source images shown in , C and D , as they were from the same experiment. D, effect of <t>SCH772984</t> on expression of ERK1/2, phosphorylated ERK1/2, and SIRT3 protein levels in KRAS/On HPNE cells. Cells were treated with SCH772984 at indicated concentrations for 24 h. Cell extracts were analyzed by Western blotting using vinculin as the loading control. HPNE KRAS/Off cell extracts were used for comparison. E, effect of SCH772984 on expression of ERK1/2, phosphorylated ERK1/2, and SIRT3 protein levels in KRAS/On HEK293 cells. Cells were treated with SCH772984 at indicated concentrations for 24 h. Cell extracts were analyzed by Western blotting using vinculin as the loading control. HEK293 KRAS/Off cell extracts were used for comparison. F, schematic illustration of the promoter activity assay to identify potential KRAS regulated region in the SIRT3 promoter. G, activity of truncated SIRT3 promoters in HPNE KRAS/Off and HPNE KRAS/On (12 h) cells. The data are presented as mean ± SD ( n = 3). p values were determined by Student’s t test. ∗, p < 0.05;∗∗, p < 0.01. HPNE: human pancreatic normal epithelial cell.
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anti-phosphorylated erk (thr202/tyr204)  (Cell Signaling Technology Inc)
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SIRT3 is transcriptionally down-regulated by KRAS G12D . A, differential gene expression of nicotinate and nicotinamide metabolism pathways. B, SIRT3 mRNA levels of HPNE cells with KRAS/Off or KRAS/On for 3 h to 96 h. The data are presented as mean ± SD ( n = 3). p values were determined by Student’s t test. ∗∗∗, p < 0.001; ∗∗∗∗, p < 0.0001. C, SIRT3 and KRAS protein levels in HPNE cells with KRAS/Off or KRAS/On for 3 h to 96 h. Cell extracts were analyzed by Western blotting using tubulin as the loading control. Note that the images of KRAS and tubulin bands were derived from the same source images shown in , C and D , as they were from the same experiment. D, effect of <t>SCH772984</t> on expression of ERK1/2, phosphorylated ERK1/2, and SIRT3 protein levels in KRAS/On HPNE cells. Cells were treated with SCH772984 at indicated concentrations for 24 h. Cell extracts were analyzed by Western blotting using vinculin as the loading control. HPNE KRAS/Off cell extracts were used for comparison. E, effect of SCH772984 on expression of ERK1/2, phosphorylated ERK1/2, and SIRT3 protein levels in KRAS/On HEK293 cells. Cells were treated with SCH772984 at indicated concentrations for 24 h. Cell extracts were analyzed by Western blotting using vinculin as the loading control. HEK293 KRAS/Off cell extracts were used for comparison. F, schematic illustration of the promoter activity assay to identify potential KRAS regulated region in the SIRT3 promoter. G, activity of truncated SIRT3 promoters in HPNE KRAS/Off and HPNE KRAS/On (12 h) cells. The data are presented as mean ± SD ( n = 3). p values were determined by Student’s t test. ∗, p < 0.05;∗∗, p < 0.01. HPNE: human pancreatic normal epithelial cell.
Anti Phosphorylated Erk (Thr202/Tyr204), supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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SIRT3 is transcriptionally down-regulated by KRAS G12D . A, differential gene expression of nicotinate and nicotinamide metabolism pathways. B, SIRT3 mRNA levels of HPNE cells with KRAS/Off or KRAS/On for 3 h to 96 h. The data are presented as mean ± SD ( n = 3). p values were determined by Student’s t test. ∗∗∗, p < 0.001; ∗∗∗∗, p < 0.0001. C, SIRT3 and KRAS protein levels in HPNE cells with KRAS/Off or KRAS/On for 3 h to 96 h. Cell extracts were analyzed by Western blotting using tubulin as the loading control. Note that the images of KRAS and tubulin bands were derived from the same source images shown in , C and D , as they were from the same experiment. D, effect of <t>SCH772984</t> on expression of ERK1/2, phosphorylated ERK1/2, and SIRT3 protein levels in KRAS/On HPNE cells. Cells were treated with SCH772984 at indicated concentrations for 24 h. Cell extracts were analyzed by Western blotting using vinculin as the loading control. HPNE KRAS/Off cell extracts were used for comparison. E, effect of SCH772984 on expression of ERK1/2, phosphorylated ERK1/2, and SIRT3 protein levels in KRAS/On HEK293 cells. Cells were treated with SCH772984 at indicated concentrations for 24 h. Cell extracts were analyzed by Western blotting using vinculin as the loading control. HEK293 KRAS/Off cell extracts were used for comparison. F, schematic illustration of the promoter activity assay to identify potential KRAS regulated region in the SIRT3 promoter. G, activity of truncated SIRT3 promoters in HPNE KRAS/Off and HPNE KRAS/On (12 h) cells. The data are presented as mean ± SD ( n = 3). p values were determined by Student’s t test. ∗, p < 0.05;∗∗, p < 0.01. HPNE: human pancreatic normal epithelial cell.
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SIRT3 is transcriptionally down-regulated by KRAS G12D . A, differential gene expression of nicotinate and nicotinamide metabolism pathways. B, SIRT3 mRNA levels of HPNE cells with KRAS/Off or KRAS/On for 3 h to 96 h. The data are presented as mean ± SD ( n = 3). p values were determined by Student’s t test. ∗∗∗, p < 0.001; ∗∗∗∗, p < 0.0001. C, SIRT3 and KRAS protein levels in HPNE cells with KRAS/Off or KRAS/On for 3 h to 96 h. Cell extracts were analyzed by Western blotting using tubulin as the loading control. Note that the images of KRAS and tubulin bands were derived from the same source images shown in , C and D , as they were from the same experiment. D, effect of SCH772984 on expression of ERK1/2, phosphorylated ERK1/2, and SIRT3 protein levels in KRAS/On HPNE cells. Cells were treated with SCH772984 at indicated concentrations for 24 h. Cell extracts were analyzed by Western blotting using vinculin as the loading control. HPNE KRAS/Off cell extracts were used for comparison. E, effect of SCH772984 on expression of ERK1/2, phosphorylated ERK1/2, and SIRT3 protein levels in KRAS/On HEK293 cells. Cells were treated with SCH772984 at indicated concentrations for 24 h. Cell extracts were analyzed by Western blotting using vinculin as the loading control. HEK293 KRAS/Off cell extracts were used for comparison. F, schematic illustration of the promoter activity assay to identify potential KRAS regulated region in the SIRT3 promoter. G, activity of truncated SIRT3 promoters in HPNE KRAS/Off and HPNE KRAS/On (12 h) cells. The data are presented as mean ± SD ( n = 3). p values were determined by Student’s t test. ∗, p < 0.05;∗∗, p < 0.01. HPNE: human pancreatic normal epithelial cell.

Journal: The Journal of Biological Chemistry

Article Title: KRAS G12D mutation promotes pancreatic tumorigenesis by suppressing sirtuin three via the guanine nucleotide exchange factor RCC1

doi: 10.1016/j.jbc.2025.111057

Figure Lengend Snippet: SIRT3 is transcriptionally down-regulated by KRAS G12D . A, differential gene expression of nicotinate and nicotinamide metabolism pathways. B, SIRT3 mRNA levels of HPNE cells with KRAS/Off or KRAS/On for 3 h to 96 h. The data are presented as mean ± SD ( n = 3). p values were determined by Student’s t test. ∗∗∗, p < 0.001; ∗∗∗∗, p < 0.0001. C, SIRT3 and KRAS protein levels in HPNE cells with KRAS/Off or KRAS/On for 3 h to 96 h. Cell extracts were analyzed by Western blotting using tubulin as the loading control. Note that the images of KRAS and tubulin bands were derived from the same source images shown in , C and D , as they were from the same experiment. D, effect of SCH772984 on expression of ERK1/2, phosphorylated ERK1/2, and SIRT3 protein levels in KRAS/On HPNE cells. Cells were treated with SCH772984 at indicated concentrations for 24 h. Cell extracts were analyzed by Western blotting using vinculin as the loading control. HPNE KRAS/Off cell extracts were used for comparison. E, effect of SCH772984 on expression of ERK1/2, phosphorylated ERK1/2, and SIRT3 protein levels in KRAS/On HEK293 cells. Cells were treated with SCH772984 at indicated concentrations for 24 h. Cell extracts were analyzed by Western blotting using vinculin as the loading control. HEK293 KRAS/Off cell extracts were used for comparison. F, schematic illustration of the promoter activity assay to identify potential KRAS regulated region in the SIRT3 promoter. G, activity of truncated SIRT3 promoters in HPNE KRAS/Off and HPNE KRAS/On (12 h) cells. The data are presented as mean ± SD ( n = 3). p values were determined by Student’s t test. ∗, p < 0.05;∗∗, p < 0.01. HPNE: human pancreatic normal epithelial cell.

Article Snippet: The ERK phosphorylation inhibitor SCH772984 (HY-50846), MYC inhibitor MCYi361 (HY-129600) and AP-1 inhibitor T-5224 (HY-12270) were purchased from MedChemExpress.

Techniques: Gene Expression, Western Blot, Control, Derivative Assay, Expressing, Comparison, Activity Assay