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anti sirtuin type 1 sirt1  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti sirtuin type 1 sirt1
    Anti Sirtuin Type 1 Sirt1, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/anti+sirtuin+1/pm41761283-106-31-36
    Average 86 stars, based on 1 article reviews
    anti sirtuin type 1 sirt1 - by Bioz Stars, 2026-10
    86/100 stars

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    Incubation:

    Article Title: Effects of American wild ginseng and Korean cultivated wild ginseng pharmacopuncture extracts on the regulation of C2C12 myoblasts differentiation through AMPK and PI3K/Akt/mTOR signaling pathway
    Article Snippet: .. The membranes underwent incubation with primary antibodies anti-peroxisome proliferator-activated receptor-γ coactivator-1-α (PGC-1α, 1:1,000; cat. no. NBP1-04676; Bioss Antibodies), anti-mitochondrial transcription factor A (TFAM, 1:1,000; cat. no. PA5-68789; Thermo Fisher Scientific, Inc.), anti-nuclear respiratory factor-1 (NRF-1, 1:1,000; cat. no. 69432s; Cell Signaling Technology, Inc.), anti-phospho-AMPK (1:500; cat. no. 44-11509; Thermo Fisher Scientific, Inc.), anti-AMPK (1:500; cat. no. AHO1332; Thermo Fisher Scientific, Inc.), anti-Sirtuin 1 (SIRT1, 1:1,000; cat. no. 69432s; Cell Signaling Technology, Inc.), anti-myosin heavy chain (MyHC, 1:1,000; cat. no. sc-376157; Santa Cruz Biotechnology, Inc.), anti-myoblast determination protein 1 (1:1,000; MyoD; cat. no. sc-377460; Santa Cruz Biotechnology, Inc.), anti-Myostatin (1:1,000; cat. no. PA5-11936, Invitrogen), anti-Myogenin (1:1,000; cat. no. sc-377460; Santa Cruz Biotechnology, Inc.), anti-phospho-AKT (1:500; cat. no. AF887; R&D Systems, Inc.), anti-AKT (1:500; cat. no. 9272s; Cell Signaling Technology, Inc.), anti-mTOR (1:500; cat. no. 29725; Cell Signaling Technology, Inc.) and anti-β-Actin (1:1,000; MilliporeSigma) overnight at 4°C. .. The membranes then underwent incubation at RT for 1 h with horseradish peroxidase-labeled anti-mouse immunoglobulin G (IgG, 1:1,000; cat no. sc-2005; Santa Cruz Biotechnology, Inc.).

    Article Title: Effects of American wild ginseng and Korean cultivated wild ginseng pharmacopuncture extracts on the regulation of C2C12 myoblasts differentiation through AMPK and PI3K/Akt/mTOR signaling pathway.
    Article Snippet: .. The membranes underwent incubation with primary antibodies anti‐peroxisome proliferator‐activated receptor‐γ coactivator‐1‐α (PGc‐1α, 1:1,000; cat. no. nBP1‐04676; Bioss antibodies), anti‐mitochondrial transcription factor a (TFaM, 1:1,000; cat. no. Pa5‐68789; Thermo Fisher Scientific, inc.), anti‐nuclear respiratory factor‐1 (nrF‐1, 1:1,000; cat. no. 69432s; cell Signaling Technology, inc.), anti‐phospho‐aMPK (1:500; cat. no. 44‐11509; Thermo Fisher Scientific, Inc.), anti‐AMPK (1:500; cat. no. AHO1332; Thermo Fisher Scientific, Inc.), anti‐Sirtuin 1 (SIRT1, 1:1,000; cat. no. 69432s; cell Signaling Technology, inc.), anti‐myosin heavy chain (MyHc, 1:1,000; cat. no. sc‐376157; Santa cruz Biotechnology, inc.), anti‐myoblast determination protein 1 (1:1,000; Myod; cat. no. sc‐377460; Santa cruz Biotechnology, inc.), anti‐Myostatin (1:1,000; cat. no. Pa5‐11936, invitrogen), anti‐Myogenin (1:1,000; cat. no. sc‐377460; Santa cruz Biotechnology, inc.), anti‐phospho‐aKT (1:500; cat. no. aF887; r&d Systems, inc.), anti‐aKT (1:500; cat. no. 9272s; cell Signaling Technology, inc.), anti‐mTor (1:500; cat. no. 29725; cell Signaling Technology, inc.) and anti‐β‐Actin (1:1,000; MilliporeSigma) overnight at 4 ̊C. .. The membranes then underwent incubation at rT for 1 h with horseradish peroxidase‐labeled anti‐mouse immunoglobulin G (igG, 1:1,000; cat no. sc‐2005; Santa cruz Biotechnology, inc.).

    Article Title: Exposure to dim light at night alters daily rhythms of glucose and lipid metabolism in rats.
    Article Snippet: Total proteins (40 or 80 μg) were separated by SDS-PAGE electrophoresis at 60 V for 20 min and then at 90 V for 80 min. Proteins were transferred to a nitrocellulose membrane using the vertical apparatus (Bio-Rad Laboratories, Hercules, CA, United States) at 230 mA for 75 min. Membranes were blocked by 1% fat-free milk or 5% bovine serum albumin (BSA) in TBS containing Tween 20 (TBS-T) for 1 h at room temperature. .. Subsequently, membranes were incubated with a primary antibody overnight at 4°C: anti-AMP-activated protein kinase α (2603, AMPKα; Cell Signaling Technology, Danvers, MA, United States, 1:1,000 dilution in 1% fat-free milk in TBST), anti-phospho-AMPKα (4188, Cell Signaling Technology, Danvers, MA, United States, 1:1,000 dilution in 1% fat-free milk in TBS-T), anti-sirtuin 1 (9475, Cell Signaling Technology, Danvers, MA, United States, 1:500 dilution in 1% BSA in TBS-T) and anti-REV-ERBα (ab174309, Abcam, Cambridge, United Kingdom, 1:1,000 dilution in 1% BSA in TBS-T). .. After thorough washing (3 min × 10 min) in TBS-T, membranes were incubated with anti-rabbit horseradish peroxidase-conjugated Frontiers in Physiology frontiersin.org04 secondary antibody (7074, Cell Signaling Technology, Danvers, MA, United States, 1:2,000 dilution in 1% fat-free milk in TBST) for 1 h at RT.

    Article Title: Exposure to dim light at night alters daily rhythms of glucose and lipid metabolism in rats
    Article Snippet: Total proteins (40 or 80 μg) were separated by SDS-PAGE electrophoresis at 60 V for 20 min and then at 90 V for 80 min. Proteins were transferred to a nitrocellulose membrane using the vertical apparatus (Bio-Rad Laboratories, Hercules, CA, United States) at 230 mA for 75 min. Membranes were blocked by 1% fat-free milk or 5% bovine serum albumin (BSA) in TBS containing Tween 20 (TBS-T) for 1 h at room temperature. .. Subsequently, membranes were incubated with a primary antibody overnight at 4°C: anti-AMP-activated protein kinase α (2603, AMPKα; Cell Signaling Technology, Danvers, MA, United States, 1:1,000 dilution in 1% fat-free milk in TBS-T), anti-phospho-AMPKα (4188, Cell Signaling Technology, Danvers, MA, United States, 1:1,000 dilution in 1% fat-free milk in TBS-T), anti-sirtuin 1 (9475, Cell Signaling Technology, Danvers, MA, United States, 1:500 dilution in 1% BSA in TBS-T) and anti-REV-ERBα (ab174309, Abcam, Cambridge, United Kingdom, 1:1,000 dilution in 1% BSA in TBS-T). .. After thorough washing (3 min × 10 min) in TBS-T, membranes were incubated with anti-rabbit horseradish peroxidase-conjugated secondary antibody (7074, Cell Signaling Technology, Danvers, MA, United States, 1:2,000 dilution in 1% fat-free milk in TBS-T) for 1 h at RT.

    Article Title: Elevation of p53 sensitizes obese kidney to adriamycin-induced aberrant lipid homeostasis via repressing HNF4α-mediated FGF21 sensitivity.
    Article Snippet: Proteins were separated by electrophoresis through 10% or 14% SDS-PAGE gels and transferred onto nitrocellulose membranes (GE Healthcare, Amersham, UK). .. The membranes were incubated with anti-NRF2 (1:1000, Abcam), anti-Kelch-like ECH-associated protein 1 (KEAP1), anti-HO-1 (1:1000, Proteintech), anti-NQO1 (1:1000, Santa Cruz), antiSREBP-1c (1:1000, Affinity Biosciences, Cincinnati, OH, USA), anti-PPARα (1:1000, Affinity Biosciences), anti-p53 (1:1000, Proteintech), anti-FGF21 (1:1000, Abcam), anti-phosphorylated extracellular regulated protein kinases (p-ERK, 1:1000, Cell Signaling Technology), anti-ERK (1:1000, Cell Signaling Technology), anti-FGFR1 (1:1000, Cell Signaling Technology), anti-p-FGFR1 (1:1000, Abcam), anti-β-Klotho (KLB, 1:1000, ABclonal Technology), anti-hepatocyte nuclear factor alpha (HNF4α, 1:1000, Abcam), anti-sirtuin 1 (SIRT1, 1:1000, Cell Signaling Technology), antiphosphorylated AMPK (1:1000, Cell Signaling Technology), anti-AMPK (1:1000, Cell Signaling Technology), anti-β-Actin (1:1000, Servicebio), anti-Na+/K+-ATPase 6 (1:1000, Cell Signaling Technology) and anti-Histone H3 (1:1000, Affinity Biosciences). .. Western blot pictures were directly taken by ImageQuant 800 (GE Healthcare).

    Article Title: MicroRNA-874-3p promotes testosterone-induced granulosa cell apoptosis by suppressing HDAC1-mediated p53 deacetylation
    Article Snippet: Proteins were separated via 10% SDS-PAGE and transferred to PVDF membranes (EMD Millipore). .. Following blocking with 5% non-fat milk at room temperature for 1 h, the membranes were incubated at 4 ̊C overnight with the following primary antibodies: Anti-androgen receptor (AR; cat. no. 5153; 1:1,000; Cell Signaling Technology, Inc.); Anti-p53 (cat. no. ab26; 1:1,000; Abcam), anti-acetylated p53 (Lys379; cat. no. 2570; 1:1,000; Cell Signaling Technology, Inc.), anti-HDAC1 (cat. no. 34589; 1:1,000; Cell Signaling Technology, Inc.), anti-sirtuin 1 (SIRT1; cat. no. 8469; 1:1,000; Cell Signaling Technology, Inc.) and anti-GAPDH (cat. no. ab8245; 1:1,000; Abcam). .. After washing by TBS with 0.05% Tween-20, the membranes were incubated with appropriate horseradish peroxidase-conjugated secondary antibodies (cat. no. ab7090 and ab97040; 1:5,000; Abcam) at room temperature for 1 h. Protein bands were visualized using an ECL kit (Beyotime Institute of Biotechnology).

    Article Title: Gastrodiae rhizoma attenuates brain aging via promoting neuritogenesis and neurodifferentiation.
    Article Snippet: Background: Gastrodiae Rhizoma (Tianma), the dried tuber of Gastrodia elata Bl. (Orchidaceae), is listed as a topgrade herbal medicine in Shen-nong Ben-ts’ao Jing and has been used for treating headaches, dizziness, vertigo and convulsion.. It has a neuroprotective effect and extends the lifespan in mouse models of Huntington’s disease and Niemann-Pick type C disease.. However, its effect on senescence remains unknown.

    Article Title: Elevation of p53 sensitizes obese kidney to adriamycin-induced aberrant lipid homeostasis via repressing HNF4α-mediated FGF21 sensitivity
    Article Snippet: Proteins were separated by electrophoresis through 10 % or 14 % SDS-PAGE gels and transferred onto nitrocellulose membranes (GE Healthcare, Amersham, UK). .. The membranes were incubated with anti-NRF2 (1:1000, Abcam), anti-Kelch-like ECH-associated protein 1 (KEAP1), anti-HO-1 (1:1000, Proteintech), anti-NQO1 (1:1000, Santa Cruz), anti-SREBP-1c (1:1000, Affinity Biosciences, Cincinnati, OH, USA), anti-PPARα (1:1000, Affinity Biosciences), anti-p53 (1:1000, Proteintech), anti-FGF21 (1:1000, Abcam), anti-phosphorylated extracellular regulated protein kinases (p-ERK, 1:1000, Cell Signaling Technology), anti-ERK (1:1000, Cell Signaling Technology), anti-FGFR1 (1:1000, Cell Signaling Technology), anti-p-FGFR1 (1:1000, Abcam), anti-β-Klotho (KLB, 1:1000, ABclonal Technology), anti-hepatocyte nuclear factor 4 alpha (HNF4α, 1:1000, Abcam), anti-sirtuin 1 (SIRT1, 1:1000, Cell Signaling Technology), anti-phosphorylated AMPK (1:1000, Cell Signaling Technology), anti-AMPK (1:1000, Cell Signaling Technology), anti-β-Actin (1:1000, Servicebio), anti-Na + /K + -ATPase (1:1000, Cell Signaling Technology) and anti-Histone H3 (1:1000, Affinity Biosciences). .. Western blot pictures were directly taken by ImageQuant 800 (GE Healthcare).

    Pyrolysis Gas Chromatography:

    Article Title: Effects of American wild ginseng and Korean cultivated wild ginseng pharmacopuncture extracts on the regulation of C2C12 myoblasts differentiation through AMPK and PI3K/Akt/mTOR signaling pathway
    Article Snippet: .. The membranes underwent incubation with primary antibodies anti-peroxisome proliferator-activated receptor-γ coactivator-1-α (PGC-1α, 1:1,000; cat. no. NBP1-04676; Bioss Antibodies), anti-mitochondrial transcription factor A (TFAM, 1:1,000; cat. no. PA5-68789; Thermo Fisher Scientific, Inc.), anti-nuclear respiratory factor-1 (NRF-1, 1:1,000; cat. no. 69432s; Cell Signaling Technology, Inc.), anti-phospho-AMPK (1:500; cat. no. 44-11509; Thermo Fisher Scientific, Inc.), anti-AMPK (1:500; cat. no. AHO1332; Thermo Fisher Scientific, Inc.), anti-Sirtuin 1 (SIRT1, 1:1,000; cat. no. 69432s; Cell Signaling Technology, Inc.), anti-myosin heavy chain (MyHC, 1:1,000; cat. no. sc-376157; Santa Cruz Biotechnology, Inc.), anti-myoblast determination protein 1 (1:1,000; MyoD; cat. no. sc-377460; Santa Cruz Biotechnology, Inc.), anti-Myostatin (1:1,000; cat. no. PA5-11936, Invitrogen), anti-Myogenin (1:1,000; cat. no. sc-377460; Santa Cruz Biotechnology, Inc.), anti-phospho-AKT (1:500; cat. no. AF887; R&D Systems, Inc.), anti-AKT (1:500; cat. no. 9272s; Cell Signaling Technology, Inc.), anti-mTOR (1:500; cat. no. 29725; Cell Signaling Technology, Inc.) and anti-β-Actin (1:1,000; MilliporeSigma) overnight at 4°C. .. The membranes then underwent incubation at RT for 1 h with horseradish peroxidase-labeled anti-mouse immunoglobulin G (IgG, 1:1,000; cat no. sc-2005; Santa Cruz Biotechnology, Inc.).

    Blocking Assay:

    Article Title: MicroRNA-874-3p promotes testosterone-induced granulosa cell apoptosis by suppressing HDAC1-mediated p53 deacetylation
    Article Snippet: Proteins were separated via 10% SDS-PAGE and transferred to PVDF membranes (EMD Millipore). .. Following blocking with 5% non-fat milk at room temperature for 1 h, the membranes were incubated at 4 ̊C overnight with the following primary antibodies: Anti-androgen receptor (AR; cat. no. 5153; 1:1,000; Cell Signaling Technology, Inc.); Anti-p53 (cat. no. ab26; 1:1,000; Abcam), anti-acetylated p53 (Lys379; cat. no. 2570; 1:1,000; Cell Signaling Technology, Inc.), anti-HDAC1 (cat. no. 34589; 1:1,000; Cell Signaling Technology, Inc.), anti-sirtuin 1 (SIRT1; cat. no. 8469; 1:1,000; Cell Signaling Technology, Inc.) and anti-GAPDH (cat. no. ab8245; 1:1,000; Abcam). .. After washing by TBS with 0.05% Tween-20, the membranes were incubated with appropriate horseradish peroxidase-conjugated secondary antibodies (cat. no. ab7090 and ab97040; 1:5,000; Abcam) at room temperature for 1 h. Protein bands were visualized using an ECL kit (Beyotime Institute of Biotechnology).



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    Decreased <t>SirT1</t> deacetylation of HSF1 mediates downregulation of heat shock proteins after EHMT2/G9a inhibition in meningioma. (A) Representative heat shock protein mRNA expression in Ben-Men I, IOMM-Lee, and CH-157MN meningioma cells after exposure to UNC0631 (4 µM) or vehicle control. Data shown are the mean ± SEM of 3 replicates for each condition. Significance was calculated using the unpaired t -test. ** P < .01, *** P < .001; ns , non-significant. (B) Western blot analysis showing downregulation of HSPA5 and SirT1 protein expression in Ben-Men-1, CH-157MN, and IOMM-Lee meningioma cells after exposure to UNC0631 (4 µM). (C) Immunoprecipitation using anti-HSF1 or control rabbit IgG antibodies followed by Western Blot using anti-HSF1 or anti-acetylated lysine antibodies. Protein lysates were obtained from CH-157MN meningioma cells exposed to UNC0631 or DMSO vehicle control. 50µg of input protein was used. (Left) input protein; (middle) IgG antibody immunoprecipitated protein; (right) HSF1 antibody immunoprecipitated protein. Arrow identifies band corresponding to acetylated HSF1. (D) Heat map of HSF1 CUT&Tag analysis in meningioma cells showing global decrease in HSF1 DNA binding (indicated in blue) at transcriptional start sites after EHMT2/G9a inhibition using UNC0631. TSS, Transcription Site Start; TES, Transcription End Site. (E) CUT&Tag analyses in IOMM-Lee cells comparing the change in HSF1 and H3K27ac signals after EHMT2/G9a inhibition with baseline EHMT2/G9a signal at cis-regulatory elements associated with genes in the apoptosis (upper panel) or UPR (lower panel) gene ontology categories. At the bottom of each heatmap, we show the “concordant regulatory elements” within each ontology that display a significant change in H3K27ac after EHMT2 inhibition along with a change in HSF1 binding to DNA or, alternatively, binding by EHMT2/G9a in untreated cells. (F) HSF1 and H3K27ac CUT&Tag analysis of promoters for genes encoding heat shock-related proteins in Ben-Men-1 and IOMM-Lee meningioma cells after exposure to UNC0631 (4 µM) or vehicle. Note the decreased binding of HSF1 to heat shock protein promoters after EHMT2/G9a inhibition. Created in BioRender. Berry (2026) https://BioRender.com/2lxmrg4 .
    Anti Sirtuin 1 Sirt1, supplied by Santa Cruz Biotechnology, 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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    Decreased <t>SirT1</t> deacetylation of HSF1 mediates downregulation of heat shock proteins after EHMT2/G9a inhibition in meningioma. (A) Representative heat shock protein mRNA expression in Ben-Men I, IOMM-Lee, and CH-157MN meningioma cells after exposure to UNC0631 (4 µM) or vehicle control. Data shown are the mean ± SEM of 3 replicates for each condition. Significance was calculated using the unpaired t -test. ** P < .01, *** P < .001; ns , non-significant. (B) Western blot analysis showing downregulation of HSPA5 and SirT1 protein expression in Ben-Men-1, CH-157MN, and IOMM-Lee meningioma cells after exposure to UNC0631 (4 µM). (C) Immunoprecipitation using anti-HSF1 or control rabbit IgG antibodies followed by Western Blot using anti-HSF1 or anti-acetylated lysine antibodies. Protein lysates were obtained from CH-157MN meningioma cells exposed to UNC0631 or DMSO vehicle control. 50µg of input protein was used. (Left) input protein; (middle) IgG antibody immunoprecipitated protein; (right) HSF1 antibody immunoprecipitated protein. Arrow identifies band corresponding to acetylated HSF1. (D) Heat map of HSF1 CUT&Tag analysis in meningioma cells showing global decrease in HSF1 DNA binding (indicated in blue) at transcriptional start sites after EHMT2/G9a inhibition using UNC0631. TSS, Transcription Site Start; TES, Transcription End Site. (E) CUT&Tag analyses in IOMM-Lee cells comparing the change in HSF1 and H3K27ac signals after EHMT2/G9a inhibition with baseline EHMT2/G9a signal at cis-regulatory elements associated with genes in the apoptosis (upper panel) or UPR (lower panel) gene ontology categories. At the bottom of each heatmap, we show the “concordant regulatory elements” within each ontology that display a significant change in H3K27ac after EHMT2 inhibition along with a change in HSF1 binding to DNA or, alternatively, binding by EHMT2/G9a in untreated cells. (F) HSF1 and H3K27ac CUT&Tag analysis of promoters for genes encoding heat shock-related proteins in Ben-Men-1 and IOMM-Lee meningioma cells after exposure to UNC0631 (4 µM) or vehicle. Note the decreased binding of HSF1 to heat shock protein promoters after EHMT2/G9a inhibition. Created in BioRender. Berry (2026) https://BioRender.com/2lxmrg4 .
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    Image Search Results


    Decreased SirT1 deacetylation of HSF1 mediates downregulation of heat shock proteins after EHMT2/G9a inhibition in meningioma. (A) Representative heat shock protein mRNA expression in Ben-Men I, IOMM-Lee, and CH-157MN meningioma cells after exposure to UNC0631 (4 µM) or vehicle control. Data shown are the mean ± SEM of 3 replicates for each condition. Significance was calculated using the unpaired t -test. ** P < .01, *** P < .001; ns , non-significant. (B) Western blot analysis showing downregulation of HSPA5 and SirT1 protein expression in Ben-Men-1, CH-157MN, and IOMM-Lee meningioma cells after exposure to UNC0631 (4 µM). (C) Immunoprecipitation using anti-HSF1 or control rabbit IgG antibodies followed by Western Blot using anti-HSF1 or anti-acetylated lysine antibodies. Protein lysates were obtained from CH-157MN meningioma cells exposed to UNC0631 or DMSO vehicle control. 50µg of input protein was used. (Left) input protein; (middle) IgG antibody immunoprecipitated protein; (right) HSF1 antibody immunoprecipitated protein. Arrow identifies band corresponding to acetylated HSF1. (D) Heat map of HSF1 CUT&Tag analysis in meningioma cells showing global decrease in HSF1 DNA binding (indicated in blue) at transcriptional start sites after EHMT2/G9a inhibition using UNC0631. TSS, Transcription Site Start; TES, Transcription End Site. (E) CUT&Tag analyses in IOMM-Lee cells comparing the change in HSF1 and H3K27ac signals after EHMT2/G9a inhibition with baseline EHMT2/G9a signal at cis-regulatory elements associated with genes in the apoptosis (upper panel) or UPR (lower panel) gene ontology categories. At the bottom of each heatmap, we show the “concordant regulatory elements” within each ontology that display a significant change in H3K27ac after EHMT2 inhibition along with a change in HSF1 binding to DNA or, alternatively, binding by EHMT2/G9a in untreated cells. (F) HSF1 and H3K27ac CUT&Tag analysis of promoters for genes encoding heat shock-related proteins in Ben-Men-1 and IOMM-Lee meningioma cells after exposure to UNC0631 (4 µM) or vehicle. Note the decreased binding of HSF1 to heat shock protein promoters after EHMT2/G9a inhibition. Created in BioRender. Berry (2026) https://BioRender.com/2lxmrg4 .

    Journal: Neuro-Oncology Advances

    Article Title: Epigenetic buffering of proteotoxic stress by EHMT2 enables meningioma growth

    doi: 10.1093/noajnl/vdag037

    Figure Lengend Snippet: Decreased SirT1 deacetylation of HSF1 mediates downregulation of heat shock proteins after EHMT2/G9a inhibition in meningioma. (A) Representative heat shock protein mRNA expression in Ben-Men I, IOMM-Lee, and CH-157MN meningioma cells after exposure to UNC0631 (4 µM) or vehicle control. Data shown are the mean ± SEM of 3 replicates for each condition. Significance was calculated using the unpaired t -test. ** P < .01, *** P < .001; ns , non-significant. (B) Western blot analysis showing downregulation of HSPA5 and SirT1 protein expression in Ben-Men-1, CH-157MN, and IOMM-Lee meningioma cells after exposure to UNC0631 (4 µM). (C) Immunoprecipitation using anti-HSF1 or control rabbit IgG antibodies followed by Western Blot using anti-HSF1 or anti-acetylated lysine antibodies. Protein lysates were obtained from CH-157MN meningioma cells exposed to UNC0631 or DMSO vehicle control. 50µg of input protein was used. (Left) input protein; (middle) IgG antibody immunoprecipitated protein; (right) HSF1 antibody immunoprecipitated protein. Arrow identifies band corresponding to acetylated HSF1. (D) Heat map of HSF1 CUT&Tag analysis in meningioma cells showing global decrease in HSF1 DNA binding (indicated in blue) at transcriptional start sites after EHMT2/G9a inhibition using UNC0631. TSS, Transcription Site Start; TES, Transcription End Site. (E) CUT&Tag analyses in IOMM-Lee cells comparing the change in HSF1 and H3K27ac signals after EHMT2/G9a inhibition with baseline EHMT2/G9a signal at cis-regulatory elements associated with genes in the apoptosis (upper panel) or UPR (lower panel) gene ontology categories. At the bottom of each heatmap, we show the “concordant regulatory elements” within each ontology that display a significant change in H3K27ac after EHMT2 inhibition along with a change in HSF1 binding to DNA or, alternatively, binding by EHMT2/G9a in untreated cells. (F) HSF1 and H3K27ac CUT&Tag analysis of promoters for genes encoding heat shock-related proteins in Ben-Men-1 and IOMM-Lee meningioma cells after exposure to UNC0631 (4 µM) or vehicle. Note the decreased binding of HSF1 to heat shock protein promoters after EHMT2/G9a inhibition. Created in BioRender. Berry (2026) https://BioRender.com/2lxmrg4 .

    Article Snippet: The following antibodies were used: anti-ATF-4 (11815, 1:1000, Cell Signaling Technology), anti-HSPA5/Bip (3177, 1:1000, Cell Signaling Technology), anti- DDIT3 /CHOP (2895, 1:1000, Cell Signaling Technology), anti-GADD34 (41 222, 1:1000, Cell Signaling Technology), anti-Sirtuin 1 (SirT1) (sc-74504, 1:1000, Santa Cruz Biotechnology), anti-HSP70 (4873, 1:1000, Cell Signaling Technology), CDKN1A /P21 (2947, 1:1000, Cell Signaling Technology), anti- BBC3 /PUMA (12450, 1:1000, Cell Signaling Technology), anti-BCL2 (2876, 1:1000, Cell Signaling Technology), anti-GAPDH (2118, 1:1000, Cell Signaling Technology), and anti-β-Actin (4970, 1:1000, Cell Signaling Technology), as well as anti-rabbit IgG, anti-mouse IgG, and anti-rat IgG secondary antibodies.

    Techniques: Inhibition, Expressing, Control, Western Blot, Immunoprecipitation, Binding Assay

    Restoration of protein folding capacity improves meningioma cell survival after EHMT2/G9a inhibition. (A) Western Blot validation of lentivirus-mediated SirT1 overexpression in meningioma cell lines compared to GFP-overexpressing control meningioma cells. (B) CCK8 cell viability assay of GFP-overexpressing (control) or SirT1-overexpressing Ben-Men-1, CH-157MN, and IOMM-Lee meningioma cells. Data shown are the mean ± SEM of 9 replicates for each condition. Statistical significance was calculated using the unpaired t -test. * P < .05; *** P < .001; ns , non-significant. (C) Western Blot analysis of HSP70 protein expression in GFP-overexpressing (control) or SirT1-overexpressing CH-157MN cells before or after EHMT2/G9a inhibition using UNC0631 (4 µM). Numbers above each band indicate normalized density of top band relative to β-actin control. (D) Fluorescence micrographs showing LIVE/DEAD cell assay of Ben-Men-1 and IOMM-Lee meningioma cells pre-incubated in 4-PBA (2 mM) or DMSO vehicle control for 2 h, followed by exposure to UNC0631 (4 µM) or DMSO control. Red color identifies dead cells, while green color identifies living cells. Scale Bar in D: 100µm. Data are quantified in bar graphs. * P < .05; ** P < .01.

    Journal: Neuro-Oncology Advances

    Article Title: Epigenetic buffering of proteotoxic stress by EHMT2 enables meningioma growth

    doi: 10.1093/noajnl/vdag037

    Figure Lengend Snippet: Restoration of protein folding capacity improves meningioma cell survival after EHMT2/G9a inhibition. (A) Western Blot validation of lentivirus-mediated SirT1 overexpression in meningioma cell lines compared to GFP-overexpressing control meningioma cells. (B) CCK8 cell viability assay of GFP-overexpressing (control) or SirT1-overexpressing Ben-Men-1, CH-157MN, and IOMM-Lee meningioma cells. Data shown are the mean ± SEM of 9 replicates for each condition. Statistical significance was calculated using the unpaired t -test. * P < .05; *** P < .001; ns , non-significant. (C) Western Blot analysis of HSP70 protein expression in GFP-overexpressing (control) or SirT1-overexpressing CH-157MN cells before or after EHMT2/G9a inhibition using UNC0631 (4 µM). Numbers above each band indicate normalized density of top band relative to β-actin control. (D) Fluorescence micrographs showing LIVE/DEAD cell assay of Ben-Men-1 and IOMM-Lee meningioma cells pre-incubated in 4-PBA (2 mM) or DMSO vehicle control for 2 h, followed by exposure to UNC0631 (4 µM) or DMSO control. Red color identifies dead cells, while green color identifies living cells. Scale Bar in D: 100µm. Data are quantified in bar graphs. * P < .05; ** P < .01.

    Article Snippet: The following antibodies were used: anti-ATF-4 (11815, 1:1000, Cell Signaling Technology), anti-HSPA5/Bip (3177, 1:1000, Cell Signaling Technology), anti- DDIT3 /CHOP (2895, 1:1000, Cell Signaling Technology), anti-GADD34 (41 222, 1:1000, Cell Signaling Technology), anti-Sirtuin 1 (SirT1) (sc-74504, 1:1000, Santa Cruz Biotechnology), anti-HSP70 (4873, 1:1000, Cell Signaling Technology), CDKN1A /P21 (2947, 1:1000, Cell Signaling Technology), anti- BBC3 /PUMA (12450, 1:1000, Cell Signaling Technology), anti-BCL2 (2876, 1:1000, Cell Signaling Technology), anti-GAPDH (2118, 1:1000, Cell Signaling Technology), and anti-β-Actin (4970, 1:1000, Cell Signaling Technology), as well as anti-rabbit IgG, anti-mouse IgG, and anti-rat IgG secondary antibodies.

    Techniques: Inhibition, Western Blot, Biomarker Discovery, Over Expression, Control, Viability Assay, Expressing, Fluorescence, Incubation