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sumo 2 3  (Proteintech)


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    Structured Review

    Proteintech sumo 2 3
    Sumo 2 3, supplied by Proteintech, used in various techniques. Bioz Stars score: 94/100, based on 27 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/sumo+2+3/SUMO2%2F3+Antibody/10__34133_slash_research__1243-286-17-19
    Average 94 stars, based on 27 article reviews
    sumo 2 3 - by Bioz Stars, 2026-09
    94/100 stars

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    Related Articles

    Western Blot:

    Article Title: YY1 enhances HIF-1α stability in tumor-associated macrophages to suppress anti-tumor immunity of prostate cancer in mice
    Article Snippet: .. Novel plant genotypes Seed stocks Authentication Plants YY1 (Abcam, cat. ab109237, 1:1000 for western blot (WB), 1:200 for immunofluorescent (IF) staining); YY1 (Boster, cat. PB9909, 1:100 for IMC); YY1 (H-10) (Santa Cruz, cat. sc-7341, 1:50 for immunoprecipitation (IP), 1:200 for IF); NUSAP1 (Proteintech, cat. 12024-1-AP, 1:2000 for WB, 1:100 for IP, 1:200 for IF); HIF-1 (abcam, cat. ab51608, 1:1000 for WB, 1:500 for IF, 1:100 for IP; cat. ab210073, 1:150 for IMC); Lyn (Proteintech, cat. 18135-1-AP, 1:1000 for WB); and Tubulin (Proteintech, cat. 11224-1-ap, 1/2000 for WB); SUMO 2/3 (Proteintech, cat. 11251-1-ap, 1:1000 for WB); Ubiquitin (Proteintech, cat. 10201-2-ap, 1:1000 for WB); Flag (Cell signaling Technology, cat. 14793S, 1:1000 for WB, 1:200 for IP); EGFP (Proteintech, cat. 66002-1-lg, 1:2000 for WB, 1:200 for IP); HA (Proteintech, cat. 51064-2-ap, 1:2000 for WB, 1:200 for IP); Phospho-Serine (Immunechem, cat. ICP9806, 1:250 for WB); PhosphoTyrosine (Immunechem, cat. ICP9805, 1:250 for WB); Phospho-YY1 (Affinity, cat. AF3694, 1:1000 for WB); GAPDH (Acton, cat. 10RG109A, 1:1000 for WB); F4/80 (Aifang biological, cat. SAF002, 1:500 for mIHC); Goat anti-mouse IgG-HRP (Santa Cruz, cat. sc2005,1:5000 for WB); CD68 (Biolegend, cat. 916104, 1:100 for IMC); Goat anti-rabbit IgG-HRP (Santa Cruz, cat. sc-2004, 1:5000 for WB); PE-CD8 (BD Bioscience, cat. 553030, 1:50 for flow cytometry (FC)); APC-CD3 (BD Bioscience, cat. 555275, 1:50 for FC); PE-CD274 (Biolegend, cat. 329706, 1:50 for FC); anti-mouse PD-1 (Bio X Cell, cat. 857122D1); Anti-mouse lgG (Bio X Cell, cat. 84932331); See supplier pages for validation. .. Abcam: https://www.abcam.cn/nav/primary-antibodies/monoclonals; Santa Cruz: https:// www.scbt.com/zh/home; Proteintech: https://www.ptgcn.com/; Cell signaling Technology: https://www.cellsignal.cn/browse/; (Immunechem: https://immunechem.com/; Affinity: https://www.affbiotech.cn/; BD Bioscience: https://www.bdbiosciences.com/zhcn/products/reagents/flow-cytometry-reagents; Bio X Cell: https://bioxcell.com/; Biolegend: https://www.biolegend.com/en-gb.

    Article Title: YY1 Enhances the Stability of HIF-1α Protein by Interacting with NUSAP1 in Macrophages within the Prostate Cancer Microenvironment
    Article Snippet: .. Reagents and antibodies used in this study include: YY1 (Abcam, cat. ab109237, 1:1000 for western blot (WB), 1:200 for immunofluorescent (IF) staining); YY1 (H-10) (Santa Cruz, cat. sc-7341, 1:50 for immunoprecipitation (IP), 1:200 for IF); NUSAP1 (Proteintech, cat. 12024-1-AP, 1:2000 for WB, 1:100 for IP, 1:200 for IF); HIF-1α (abcam, cat. ab51608, 1:1000 for WB, 1:500 for IF, 1:100 for IP); Lyn (Proteintech, cat. 18135-1-AP, 1:1000 for WB); and Tubulin (Proteintech, cat. 11224-1-ap, 1/2000 for WB); SUMO 2/3 (Proteintech, cat. 11251-1-ap, 1:1000 for WB); Ubiquitin (Proteintech, cat. 10201-2-ap, 1:1000 for WB); Flag (Cell signaling Technology, cat. 14793S, 1:1000 for WB, 1:200 for IP); EGFP (Proteintech, cat. 66002-1-lg, 1:2000 for WB, 1:200 for IP); HA (Proteintech, cat. 51064-2-ap, 1:2000 for WB, 1:200 for IP); Phospho-Serine (Immunechem, cat. ICP9806, 1:250 for WB); Phospho-Tyrosine (Immunechem, cat. ICP9805, 1:250 for WB); Phospho-YY1 (Affinity, cat. AF3694, 1:1000 for WB); GAPDH (Acton, cat. 10R-G109A, 1:1000 for WB); F4/80 (Aifang biological, cat. SAF002, 1:500 for mIHC); Goat anti-mouse IgG-HRP (Santa Cruz, cat. sc2005,1:5000 for WB); Goat anti-rabbit IgG-HRP (Santa Cruz, cat. sc-2004, 1:5000 for WB); PE-CD8 (BD Bioscience, cat. 553030, 1:50 for flow cytometry (FC)); APC-CD3 (BD Bioscience, cat. 555275, 1:50 for FC); PE-CD274 (Biolegend, cat. 329706, 1:50 for FC); 1,6-hexanediol (Aladdin, cat. H103708); Tenapanor (TargetMol, T7587, cat. 1234423-95-0); MG132 (MACKLIN, cat. HY132-59); CHX (MACKLIN, cat. HY-12320, 100 μg/ml for CHX assay); Subasumstat (MACKLIN, TAK-981, cat. HY-111789); Na 3 VO 4 (MACKLIN, cat. 13721-39-6); SU6656 (MACKLIN, cat. HY-B078); K3PO4 (MACKLIN, cat. NONE8222); Protein A/G Magnetic Beads (Beyotime, cat. P2108); Anlotinib (cat. AL3818) dihydrochloride (Aladdin, cat. 1360460-82-7); Cobalt Chloride Solution (MACKLIN, cat. 7791-13-1); anti-mouse PD-1 (Bio X Cell, cat. 857122D1); Anti-mouse lgG (Bio X Cell, cat. 84932331); The full-length coding sequences of YY1, HIF-1α and NUSAP1 (or their variants), as well as IDRs of YY1 and HIF-1α were individually subcloned into a modiffed version of a pGEX vector with 6 × His and EGFP or mCherry at the N-terminus. ..

    Staining:

    Article Title: YY1 enhances HIF-1α stability in tumor-associated macrophages to suppress anti-tumor immunity of prostate cancer in mice
    Article Snippet: .. Novel plant genotypes Seed stocks Authentication Plants YY1 (Abcam, cat. ab109237, 1:1000 for western blot (WB), 1:200 for immunofluorescent (IF) staining); YY1 (Boster, cat. PB9909, 1:100 for IMC); YY1 (H-10) (Santa Cruz, cat. sc-7341, 1:50 for immunoprecipitation (IP), 1:200 for IF); NUSAP1 (Proteintech, cat. 12024-1-AP, 1:2000 for WB, 1:100 for IP, 1:200 for IF); HIF-1 (abcam, cat. ab51608, 1:1000 for WB, 1:500 for IF, 1:100 for IP; cat. ab210073, 1:150 for IMC); Lyn (Proteintech, cat. 18135-1-AP, 1:1000 for WB); and Tubulin (Proteintech, cat. 11224-1-ap, 1/2000 for WB); SUMO 2/3 (Proteintech, cat. 11251-1-ap, 1:1000 for WB); Ubiquitin (Proteintech, cat. 10201-2-ap, 1:1000 for WB); Flag (Cell signaling Technology, cat. 14793S, 1:1000 for WB, 1:200 for IP); EGFP (Proteintech, cat. 66002-1-lg, 1:2000 for WB, 1:200 for IP); HA (Proteintech, cat. 51064-2-ap, 1:2000 for WB, 1:200 for IP); Phospho-Serine (Immunechem, cat. ICP9806, 1:250 for WB); PhosphoTyrosine (Immunechem, cat. ICP9805, 1:250 for WB); Phospho-YY1 (Affinity, cat. AF3694, 1:1000 for WB); GAPDH (Acton, cat. 10RG109A, 1:1000 for WB); F4/80 (Aifang biological, cat. SAF002, 1:500 for mIHC); Goat anti-mouse IgG-HRP (Santa Cruz, cat. sc2005,1:5000 for WB); CD68 (Biolegend, cat. 916104, 1:100 for IMC); Goat anti-rabbit IgG-HRP (Santa Cruz, cat. sc-2004, 1:5000 for WB); PE-CD8 (BD Bioscience, cat. 553030, 1:50 for flow cytometry (FC)); APC-CD3 (BD Bioscience, cat. 555275, 1:50 for FC); PE-CD274 (Biolegend, cat. 329706, 1:50 for FC); anti-mouse PD-1 (Bio X Cell, cat. 857122D1); Anti-mouse lgG (Bio X Cell, cat. 84932331); See supplier pages for validation. .. Abcam: https://www.abcam.cn/nav/primary-antibodies/monoclonals; Santa Cruz: https:// www.scbt.com/zh/home; Proteintech: https://www.ptgcn.com/; Cell signaling Technology: https://www.cellsignal.cn/browse/; (Immunechem: https://immunechem.com/; Affinity: https://www.affbiotech.cn/; BD Bioscience: https://www.bdbiosciences.com/zhcn/products/reagents/flow-cytometry-reagents; Bio X Cell: https://bioxcell.com/; Biolegend: https://www.biolegend.com/en-gb.

    Article Title: YY1 Enhances the Stability of HIF-1α Protein by Interacting with NUSAP1 in Macrophages within the Prostate Cancer Microenvironment
    Article Snippet: .. Reagents and antibodies used in this study include: YY1 (Abcam, cat. ab109237, 1:1000 for western blot (WB), 1:200 for immunofluorescent (IF) staining); YY1 (H-10) (Santa Cruz, cat. sc-7341, 1:50 for immunoprecipitation (IP), 1:200 for IF); NUSAP1 (Proteintech, cat. 12024-1-AP, 1:2000 for WB, 1:100 for IP, 1:200 for IF); HIF-1α (abcam, cat. ab51608, 1:1000 for WB, 1:500 for IF, 1:100 for IP); Lyn (Proteintech, cat. 18135-1-AP, 1:1000 for WB); and Tubulin (Proteintech, cat. 11224-1-ap, 1/2000 for WB); SUMO 2/3 (Proteintech, cat. 11251-1-ap, 1:1000 for WB); Ubiquitin (Proteintech, cat. 10201-2-ap, 1:1000 for WB); Flag (Cell signaling Technology, cat. 14793S, 1:1000 for WB, 1:200 for IP); EGFP (Proteintech, cat. 66002-1-lg, 1:2000 for WB, 1:200 for IP); HA (Proteintech, cat. 51064-2-ap, 1:2000 for WB, 1:200 for IP); Phospho-Serine (Immunechem, cat. ICP9806, 1:250 for WB); Phospho-Tyrosine (Immunechem, cat. ICP9805, 1:250 for WB); Phospho-YY1 (Affinity, cat. AF3694, 1:1000 for WB); GAPDH (Acton, cat. 10R-G109A, 1:1000 for WB); F4/80 (Aifang biological, cat. SAF002, 1:500 for mIHC); Goat anti-mouse IgG-HRP (Santa Cruz, cat. sc2005,1:5000 for WB); Goat anti-rabbit IgG-HRP (Santa Cruz, cat. sc-2004, 1:5000 for WB); PE-CD8 (BD Bioscience, cat. 553030, 1:50 for flow cytometry (FC)); APC-CD3 (BD Bioscience, cat. 555275, 1:50 for FC); PE-CD274 (Biolegend, cat. 329706, 1:50 for FC); 1,6-hexanediol (Aladdin, cat. H103708); Tenapanor (TargetMol, T7587, cat. 1234423-95-0); MG132 (MACKLIN, cat. HY132-59); CHX (MACKLIN, cat. HY-12320, 100 μg/ml for CHX assay); Subasumstat (MACKLIN, TAK-981, cat. HY-111789); Na 3 VO 4 (MACKLIN, cat. 13721-39-6); SU6656 (MACKLIN, cat. HY-B078); K3PO4 (MACKLIN, cat. NONE8222); Protein A/G Magnetic Beads (Beyotime, cat. P2108); Anlotinib (cat. AL3818) dihydrochloride (Aladdin, cat. 1360460-82-7); Cobalt Chloride Solution (MACKLIN, cat. 7791-13-1); anti-mouse PD-1 (Bio X Cell, cat. 857122D1); Anti-mouse lgG (Bio X Cell, cat. 84932331); The full-length coding sequences of YY1, HIF-1α and NUSAP1 (or their variants), as well as IDRs of YY1 and HIF-1α were individually subcloned into a modiffed version of a pGEX vector with 6 × His and EGFP or mCherry at the N-terminus. ..

    Immunoprecipitation:

    Article Title: YY1 enhances HIF-1α stability in tumor-associated macrophages to suppress anti-tumor immunity of prostate cancer in mice
    Article Snippet: .. Novel plant genotypes Seed stocks Authentication Plants YY1 (Abcam, cat. ab109237, 1:1000 for western blot (WB), 1:200 for immunofluorescent (IF) staining); YY1 (Boster, cat. PB9909, 1:100 for IMC); YY1 (H-10) (Santa Cruz, cat. sc-7341, 1:50 for immunoprecipitation (IP), 1:200 for IF); NUSAP1 (Proteintech, cat. 12024-1-AP, 1:2000 for WB, 1:100 for IP, 1:200 for IF); HIF-1 (abcam, cat. ab51608, 1:1000 for WB, 1:500 for IF, 1:100 for IP; cat. ab210073, 1:150 for IMC); Lyn (Proteintech, cat. 18135-1-AP, 1:1000 for WB); and Tubulin (Proteintech, cat. 11224-1-ap, 1/2000 for WB); SUMO 2/3 (Proteintech, cat. 11251-1-ap, 1:1000 for WB); Ubiquitin (Proteintech, cat. 10201-2-ap, 1:1000 for WB); Flag (Cell signaling Technology, cat. 14793S, 1:1000 for WB, 1:200 for IP); EGFP (Proteintech, cat. 66002-1-lg, 1:2000 for WB, 1:200 for IP); HA (Proteintech, cat. 51064-2-ap, 1:2000 for WB, 1:200 for IP); Phospho-Serine (Immunechem, cat. ICP9806, 1:250 for WB); PhosphoTyrosine (Immunechem, cat. ICP9805, 1:250 for WB); Phospho-YY1 (Affinity, cat. AF3694, 1:1000 for WB); GAPDH (Acton, cat. 10RG109A, 1:1000 for WB); F4/80 (Aifang biological, cat. SAF002, 1:500 for mIHC); Goat anti-mouse IgG-HRP (Santa Cruz, cat. sc2005,1:5000 for WB); CD68 (Biolegend, cat. 916104, 1:100 for IMC); Goat anti-rabbit IgG-HRP (Santa Cruz, cat. sc-2004, 1:5000 for WB); PE-CD8 (BD Bioscience, cat. 553030, 1:50 for flow cytometry (FC)); APC-CD3 (BD Bioscience, cat. 555275, 1:50 for FC); PE-CD274 (Biolegend, cat. 329706, 1:50 for FC); anti-mouse PD-1 (Bio X Cell, cat. 857122D1); Anti-mouse lgG (Bio X Cell, cat. 84932331); See supplier pages for validation. .. Abcam: https://www.abcam.cn/nav/primary-antibodies/monoclonals; Santa Cruz: https:// www.scbt.com/zh/home; Proteintech: https://www.ptgcn.com/; Cell signaling Technology: https://www.cellsignal.cn/browse/; (Immunechem: https://immunechem.com/; Affinity: https://www.affbiotech.cn/; BD Bioscience: https://www.bdbiosciences.com/zhcn/products/reagents/flow-cytometry-reagents; Bio X Cell: https://bioxcell.com/; Biolegend: https://www.biolegend.com/en-gb.

    Article Title: YY1 Enhances the Stability of HIF-1α Protein by Interacting with NUSAP1 in Macrophages within the Prostate Cancer Microenvironment
    Article Snippet: .. Reagents and antibodies used in this study include: YY1 (Abcam, cat. ab109237, 1:1000 for western blot (WB), 1:200 for immunofluorescent (IF) staining); YY1 (H-10) (Santa Cruz, cat. sc-7341, 1:50 for immunoprecipitation (IP), 1:200 for IF); NUSAP1 (Proteintech, cat. 12024-1-AP, 1:2000 for WB, 1:100 for IP, 1:200 for IF); HIF-1α (abcam, cat. ab51608, 1:1000 for WB, 1:500 for IF, 1:100 for IP); Lyn (Proteintech, cat. 18135-1-AP, 1:1000 for WB); and Tubulin (Proteintech, cat. 11224-1-ap, 1/2000 for WB); SUMO 2/3 (Proteintech, cat. 11251-1-ap, 1:1000 for WB); Ubiquitin (Proteintech, cat. 10201-2-ap, 1:1000 for WB); Flag (Cell signaling Technology, cat. 14793S, 1:1000 for WB, 1:200 for IP); EGFP (Proteintech, cat. 66002-1-lg, 1:2000 for WB, 1:200 for IP); HA (Proteintech, cat. 51064-2-ap, 1:2000 for WB, 1:200 for IP); Phospho-Serine (Immunechem, cat. ICP9806, 1:250 for WB); Phospho-Tyrosine (Immunechem, cat. ICP9805, 1:250 for WB); Phospho-YY1 (Affinity, cat. AF3694, 1:1000 for WB); GAPDH (Acton, cat. 10R-G109A, 1:1000 for WB); F4/80 (Aifang biological, cat. SAF002, 1:500 for mIHC); Goat anti-mouse IgG-HRP (Santa Cruz, cat. sc2005,1:5000 for WB); Goat anti-rabbit IgG-HRP (Santa Cruz, cat. sc-2004, 1:5000 for WB); PE-CD8 (BD Bioscience, cat. 553030, 1:50 for flow cytometry (FC)); APC-CD3 (BD Bioscience, cat. 555275, 1:50 for FC); PE-CD274 (Biolegend, cat. 329706, 1:50 for FC); 1,6-hexanediol (Aladdin, cat. H103708); Tenapanor (TargetMol, T7587, cat. 1234423-95-0); MG132 (MACKLIN, cat. HY132-59); CHX (MACKLIN, cat. HY-12320, 100 μg/ml for CHX assay); Subasumstat (MACKLIN, TAK-981, cat. HY-111789); Na 3 VO 4 (MACKLIN, cat. 13721-39-6); SU6656 (MACKLIN, cat. HY-B078); K3PO4 (MACKLIN, cat. NONE8222); Protein A/G Magnetic Beads (Beyotime, cat. P2108); Anlotinib (cat. AL3818) dihydrochloride (Aladdin, cat. 1360460-82-7); Cobalt Chloride Solution (MACKLIN, cat. 7791-13-1); anti-mouse PD-1 (Bio X Cell, cat. 857122D1); Anti-mouse lgG (Bio X Cell, cat. 84932331); The full-length coding sequences of YY1, HIF-1α and NUSAP1 (or their variants), as well as IDRs of YY1 and HIF-1α were individually subcloned into a modiffed version of a pGEX vector with 6 × His and EGFP or mCherry at the N-terminus. ..

    Ubiquitin Proteomics:

    Article Title: YY1 enhances HIF-1α stability in tumor-associated macrophages to suppress anti-tumor immunity of prostate cancer in mice
    Article Snippet: .. Novel plant genotypes Seed stocks Authentication Plants YY1 (Abcam, cat. ab109237, 1:1000 for western blot (WB), 1:200 for immunofluorescent (IF) staining); YY1 (Boster, cat. PB9909, 1:100 for IMC); YY1 (H-10) (Santa Cruz, cat. sc-7341, 1:50 for immunoprecipitation (IP), 1:200 for IF); NUSAP1 (Proteintech, cat. 12024-1-AP, 1:2000 for WB, 1:100 for IP, 1:200 for IF); HIF-1 (abcam, cat. ab51608, 1:1000 for WB, 1:500 for IF, 1:100 for IP; cat. ab210073, 1:150 for IMC); Lyn (Proteintech, cat. 18135-1-AP, 1:1000 for WB); and Tubulin (Proteintech, cat. 11224-1-ap, 1/2000 for WB); SUMO 2/3 (Proteintech, cat. 11251-1-ap, 1:1000 for WB); Ubiquitin (Proteintech, cat. 10201-2-ap, 1:1000 for WB); Flag (Cell signaling Technology, cat. 14793S, 1:1000 for WB, 1:200 for IP); EGFP (Proteintech, cat. 66002-1-lg, 1:2000 for WB, 1:200 for IP); HA (Proteintech, cat. 51064-2-ap, 1:2000 for WB, 1:200 for IP); Phospho-Serine (Immunechem, cat. ICP9806, 1:250 for WB); PhosphoTyrosine (Immunechem, cat. ICP9805, 1:250 for WB); Phospho-YY1 (Affinity, cat. AF3694, 1:1000 for WB); GAPDH (Acton, cat. 10RG109A, 1:1000 for WB); F4/80 (Aifang biological, cat. SAF002, 1:500 for mIHC); Goat anti-mouse IgG-HRP (Santa Cruz, cat. sc2005,1:5000 for WB); CD68 (Biolegend, cat. 916104, 1:100 for IMC); Goat anti-rabbit IgG-HRP (Santa Cruz, cat. sc-2004, 1:5000 for WB); PE-CD8 (BD Bioscience, cat. 553030, 1:50 for flow cytometry (FC)); APC-CD3 (BD Bioscience, cat. 555275, 1:50 for FC); PE-CD274 (Biolegend, cat. 329706, 1:50 for FC); anti-mouse PD-1 (Bio X Cell, cat. 857122D1); Anti-mouse lgG (Bio X Cell, cat. 84932331); See supplier pages for validation. .. Abcam: https://www.abcam.cn/nav/primary-antibodies/monoclonals; Santa Cruz: https:// www.scbt.com/zh/home; Proteintech: https://www.ptgcn.com/; Cell signaling Technology: https://www.cellsignal.cn/browse/; (Immunechem: https://immunechem.com/; Affinity: https://www.affbiotech.cn/; BD Bioscience: https://www.bdbiosciences.com/zhcn/products/reagents/flow-cytometry-reagents; Bio X Cell: https://bioxcell.com/; Biolegend: https://www.biolegend.com/en-gb.

    Article Title: YY1 Enhances the Stability of HIF-1α Protein by Interacting with NUSAP1 in Macrophages within the Prostate Cancer Microenvironment
    Article Snippet: .. Reagents and antibodies used in this study include: YY1 (Abcam, cat. ab109237, 1:1000 for western blot (WB), 1:200 for immunofluorescent (IF) staining); YY1 (H-10) (Santa Cruz, cat. sc-7341, 1:50 for immunoprecipitation (IP), 1:200 for IF); NUSAP1 (Proteintech, cat. 12024-1-AP, 1:2000 for WB, 1:100 for IP, 1:200 for IF); HIF-1α (abcam, cat. ab51608, 1:1000 for WB, 1:500 for IF, 1:100 for IP); Lyn (Proteintech, cat. 18135-1-AP, 1:1000 for WB); and Tubulin (Proteintech, cat. 11224-1-ap, 1/2000 for WB); SUMO 2/3 (Proteintech, cat. 11251-1-ap, 1:1000 for WB); Ubiquitin (Proteintech, cat. 10201-2-ap, 1:1000 for WB); Flag (Cell signaling Technology, cat. 14793S, 1:1000 for WB, 1:200 for IP); EGFP (Proteintech, cat. 66002-1-lg, 1:2000 for WB, 1:200 for IP); HA (Proteintech, cat. 51064-2-ap, 1:2000 for WB, 1:200 for IP); Phospho-Serine (Immunechem, cat. ICP9806, 1:250 for WB); Phospho-Tyrosine (Immunechem, cat. ICP9805, 1:250 for WB); Phospho-YY1 (Affinity, cat. AF3694, 1:1000 for WB); GAPDH (Acton, cat. 10R-G109A, 1:1000 for WB); F4/80 (Aifang biological, cat. SAF002, 1:500 for mIHC); Goat anti-mouse IgG-HRP (Santa Cruz, cat. sc2005,1:5000 for WB); Goat anti-rabbit IgG-HRP (Santa Cruz, cat. sc-2004, 1:5000 for WB); PE-CD8 (BD Bioscience, cat. 553030, 1:50 for flow cytometry (FC)); APC-CD3 (BD Bioscience, cat. 555275, 1:50 for FC); PE-CD274 (Biolegend, cat. 329706, 1:50 for FC); 1,6-hexanediol (Aladdin, cat. H103708); Tenapanor (TargetMol, T7587, cat. 1234423-95-0); MG132 (MACKLIN, cat. HY132-59); CHX (MACKLIN, cat. HY-12320, 100 μg/ml for CHX assay); Subasumstat (MACKLIN, TAK-981, cat. HY-111789); Na 3 VO 4 (MACKLIN, cat. 13721-39-6); SU6656 (MACKLIN, cat. HY-B078); K3PO4 (MACKLIN, cat. NONE8222); Protein A/G Magnetic Beads (Beyotime, cat. P2108); Anlotinib (cat. AL3818) dihydrochloride (Aladdin, cat. 1360460-82-7); Cobalt Chloride Solution (MACKLIN, cat. 7791-13-1); anti-mouse PD-1 (Bio X Cell, cat. 857122D1); Anti-mouse lgG (Bio X Cell, cat. 84932331); The full-length coding sequences of YY1, HIF-1α and NUSAP1 (or their variants), as well as IDRs of YY1 and HIF-1α were individually subcloned into a modiffed version of a pGEX vector with 6 × His and EGFP or mCherry at the N-terminus. ..

    Flow Cytometry:

    Article Title: YY1 enhances HIF-1α stability in tumor-associated macrophages to suppress anti-tumor immunity of prostate cancer in mice
    Article Snippet: .. Novel plant genotypes Seed stocks Authentication Plants YY1 (Abcam, cat. ab109237, 1:1000 for western blot (WB), 1:200 for immunofluorescent (IF) staining); YY1 (Boster, cat. PB9909, 1:100 for IMC); YY1 (H-10) (Santa Cruz, cat. sc-7341, 1:50 for immunoprecipitation (IP), 1:200 for IF); NUSAP1 (Proteintech, cat. 12024-1-AP, 1:2000 for WB, 1:100 for IP, 1:200 for IF); HIF-1 (abcam, cat. ab51608, 1:1000 for WB, 1:500 for IF, 1:100 for IP; cat. ab210073, 1:150 for IMC); Lyn (Proteintech, cat. 18135-1-AP, 1:1000 for WB); and Tubulin (Proteintech, cat. 11224-1-ap, 1/2000 for WB); SUMO 2/3 (Proteintech, cat. 11251-1-ap, 1:1000 for WB); Ubiquitin (Proteintech, cat. 10201-2-ap, 1:1000 for WB); Flag (Cell signaling Technology, cat. 14793S, 1:1000 for WB, 1:200 for IP); EGFP (Proteintech, cat. 66002-1-lg, 1:2000 for WB, 1:200 for IP); HA (Proteintech, cat. 51064-2-ap, 1:2000 for WB, 1:200 for IP); Phospho-Serine (Immunechem, cat. ICP9806, 1:250 for WB); PhosphoTyrosine (Immunechem, cat. ICP9805, 1:250 for WB); Phospho-YY1 (Affinity, cat. AF3694, 1:1000 for WB); GAPDH (Acton, cat. 10RG109A, 1:1000 for WB); F4/80 (Aifang biological, cat. SAF002, 1:500 for mIHC); Goat anti-mouse IgG-HRP (Santa Cruz, cat. sc2005,1:5000 for WB); CD68 (Biolegend, cat. 916104, 1:100 for IMC); Goat anti-rabbit IgG-HRP (Santa Cruz, cat. sc-2004, 1:5000 for WB); PE-CD8 (BD Bioscience, cat. 553030, 1:50 for flow cytometry (FC)); APC-CD3 (BD Bioscience, cat. 555275, 1:50 for FC); PE-CD274 (Biolegend, cat. 329706, 1:50 for FC); anti-mouse PD-1 (Bio X Cell, cat. 857122D1); Anti-mouse lgG (Bio X Cell, cat. 84932331); See supplier pages for validation. .. Abcam: https://www.abcam.cn/nav/primary-antibodies/monoclonals; Santa Cruz: https:// www.scbt.com/zh/home; Proteintech: https://www.ptgcn.com/; Cell signaling Technology: https://www.cellsignal.cn/browse/; (Immunechem: https://immunechem.com/; Affinity: https://www.affbiotech.cn/; BD Bioscience: https://www.bdbiosciences.com/zhcn/products/reagents/flow-cytometry-reagents; Bio X Cell: https://bioxcell.com/; Biolegend: https://www.biolegend.com/en-gb.

    Article Title: YY1 Enhances the Stability of HIF-1α Protein by Interacting with NUSAP1 in Macrophages within the Prostate Cancer Microenvironment
    Article Snippet: .. Reagents and antibodies used in this study include: YY1 (Abcam, cat. ab109237, 1:1000 for western blot (WB), 1:200 for immunofluorescent (IF) staining); YY1 (H-10) (Santa Cruz, cat. sc-7341, 1:50 for immunoprecipitation (IP), 1:200 for IF); NUSAP1 (Proteintech, cat. 12024-1-AP, 1:2000 for WB, 1:100 for IP, 1:200 for IF); HIF-1α (abcam, cat. ab51608, 1:1000 for WB, 1:500 for IF, 1:100 for IP); Lyn (Proteintech, cat. 18135-1-AP, 1:1000 for WB); and Tubulin (Proteintech, cat. 11224-1-ap, 1/2000 for WB); SUMO 2/3 (Proteintech, cat. 11251-1-ap, 1:1000 for WB); Ubiquitin (Proteintech, cat. 10201-2-ap, 1:1000 for WB); Flag (Cell signaling Technology, cat. 14793S, 1:1000 for WB, 1:200 for IP); EGFP (Proteintech, cat. 66002-1-lg, 1:2000 for WB, 1:200 for IP); HA (Proteintech, cat. 51064-2-ap, 1:2000 for WB, 1:200 for IP); Phospho-Serine (Immunechem, cat. ICP9806, 1:250 for WB); Phospho-Tyrosine (Immunechem, cat. ICP9805, 1:250 for WB); Phospho-YY1 (Affinity, cat. AF3694, 1:1000 for WB); GAPDH (Acton, cat. 10R-G109A, 1:1000 for WB); F4/80 (Aifang biological, cat. SAF002, 1:500 for mIHC); Goat anti-mouse IgG-HRP (Santa Cruz, cat. sc2005,1:5000 for WB); Goat anti-rabbit IgG-HRP (Santa Cruz, cat. sc-2004, 1:5000 for WB); PE-CD8 (BD Bioscience, cat. 553030, 1:50 for flow cytometry (FC)); APC-CD3 (BD Bioscience, cat. 555275, 1:50 for FC); PE-CD274 (Biolegend, cat. 329706, 1:50 for FC); 1,6-hexanediol (Aladdin, cat. H103708); Tenapanor (TargetMol, T7587, cat. 1234423-95-0); MG132 (MACKLIN, cat. HY132-59); CHX (MACKLIN, cat. HY-12320, 100 μg/ml for CHX assay); Subasumstat (MACKLIN, TAK-981, cat. HY-111789); Na 3 VO 4 (MACKLIN, cat. 13721-39-6); SU6656 (MACKLIN, cat. HY-B078); K3PO4 (MACKLIN, cat. NONE8222); Protein A/G Magnetic Beads (Beyotime, cat. P2108); Anlotinib (cat. AL3818) dihydrochloride (Aladdin, cat. 1360460-82-7); Cobalt Chloride Solution (MACKLIN, cat. 7791-13-1); anti-mouse PD-1 (Bio X Cell, cat. 857122D1); Anti-mouse lgG (Bio X Cell, cat. 84932331); The full-length coding sequences of YY1, HIF-1α and NUSAP1 (or their variants), as well as IDRs of YY1 and HIF-1α were individually subcloned into a modiffed version of a pGEX vector with 6 × His and EGFP or mCherry at the N-terminus. ..

    Biomarker Discovery:

    Article Title: YY1 enhances HIF-1α stability in tumor-associated macrophages to suppress anti-tumor immunity of prostate cancer in mice
    Article Snippet: .. Novel plant genotypes Seed stocks Authentication Plants YY1 (Abcam, cat. ab109237, 1:1000 for western blot (WB), 1:200 for immunofluorescent (IF) staining); YY1 (Boster, cat. PB9909, 1:100 for IMC); YY1 (H-10) (Santa Cruz, cat. sc-7341, 1:50 for immunoprecipitation (IP), 1:200 for IF); NUSAP1 (Proteintech, cat. 12024-1-AP, 1:2000 for WB, 1:100 for IP, 1:200 for IF); HIF-1 (abcam, cat. ab51608, 1:1000 for WB, 1:500 for IF, 1:100 for IP; cat. ab210073, 1:150 for IMC); Lyn (Proteintech, cat. 18135-1-AP, 1:1000 for WB); and Tubulin (Proteintech, cat. 11224-1-ap, 1/2000 for WB); SUMO 2/3 (Proteintech, cat. 11251-1-ap, 1:1000 for WB); Ubiquitin (Proteintech, cat. 10201-2-ap, 1:1000 for WB); Flag (Cell signaling Technology, cat. 14793S, 1:1000 for WB, 1:200 for IP); EGFP (Proteintech, cat. 66002-1-lg, 1:2000 for WB, 1:200 for IP); HA (Proteintech, cat. 51064-2-ap, 1:2000 for WB, 1:200 for IP); Phospho-Serine (Immunechem, cat. ICP9806, 1:250 for WB); PhosphoTyrosine (Immunechem, cat. ICP9805, 1:250 for WB); Phospho-YY1 (Affinity, cat. AF3694, 1:1000 for WB); GAPDH (Acton, cat. 10RG109A, 1:1000 for WB); F4/80 (Aifang biological, cat. SAF002, 1:500 for mIHC); Goat anti-mouse IgG-HRP (Santa Cruz, cat. sc2005,1:5000 for WB); CD68 (Biolegend, cat. 916104, 1:100 for IMC); Goat anti-rabbit IgG-HRP (Santa Cruz, cat. sc-2004, 1:5000 for WB); PE-CD8 (BD Bioscience, cat. 553030, 1:50 for flow cytometry (FC)); APC-CD3 (BD Bioscience, cat. 555275, 1:50 for FC); PE-CD274 (Biolegend, cat. 329706, 1:50 for FC); anti-mouse PD-1 (Bio X Cell, cat. 857122D1); Anti-mouse lgG (Bio X Cell, cat. 84932331); See supplier pages for validation. .. Abcam: https://www.abcam.cn/nav/primary-antibodies/monoclonals; Santa Cruz: https:// www.scbt.com/zh/home; Proteintech: https://www.ptgcn.com/; Cell signaling Technology: https://www.cellsignal.cn/browse/; (Immunechem: https://immunechem.com/; Affinity: https://www.affbiotech.cn/; BD Bioscience: https://www.bdbiosciences.com/zhcn/products/reagents/flow-cytometry-reagents; Bio X Cell: https://bioxcell.com/; Biolegend: https://www.biolegend.com/en-gb.

    Magnetic Beads:

    Article Title: YY1 Enhances the Stability of HIF-1α Protein by Interacting with NUSAP1 in Macrophages within the Prostate Cancer Microenvironment
    Article Snippet: .. Reagents and antibodies used in this study include: YY1 (Abcam, cat. ab109237, 1:1000 for western blot (WB), 1:200 for immunofluorescent (IF) staining); YY1 (H-10) (Santa Cruz, cat. sc-7341, 1:50 for immunoprecipitation (IP), 1:200 for IF); NUSAP1 (Proteintech, cat. 12024-1-AP, 1:2000 for WB, 1:100 for IP, 1:200 for IF); HIF-1α (abcam, cat. ab51608, 1:1000 for WB, 1:500 for IF, 1:100 for IP); Lyn (Proteintech, cat. 18135-1-AP, 1:1000 for WB); and Tubulin (Proteintech, cat. 11224-1-ap, 1/2000 for WB); SUMO 2/3 (Proteintech, cat. 11251-1-ap, 1:1000 for WB); Ubiquitin (Proteintech, cat. 10201-2-ap, 1:1000 for WB); Flag (Cell signaling Technology, cat. 14793S, 1:1000 for WB, 1:200 for IP); EGFP (Proteintech, cat. 66002-1-lg, 1:2000 for WB, 1:200 for IP); HA (Proteintech, cat. 51064-2-ap, 1:2000 for WB, 1:200 for IP); Phospho-Serine (Immunechem, cat. ICP9806, 1:250 for WB); Phospho-Tyrosine (Immunechem, cat. ICP9805, 1:250 for WB); Phospho-YY1 (Affinity, cat. AF3694, 1:1000 for WB); GAPDH (Acton, cat. 10R-G109A, 1:1000 for WB); F4/80 (Aifang biological, cat. SAF002, 1:500 for mIHC); Goat anti-mouse IgG-HRP (Santa Cruz, cat. sc2005,1:5000 for WB); Goat anti-rabbit IgG-HRP (Santa Cruz, cat. sc-2004, 1:5000 for WB); PE-CD8 (BD Bioscience, cat. 553030, 1:50 for flow cytometry (FC)); APC-CD3 (BD Bioscience, cat. 555275, 1:50 for FC); PE-CD274 (Biolegend, cat. 329706, 1:50 for FC); 1,6-hexanediol (Aladdin, cat. H103708); Tenapanor (TargetMol, T7587, cat. 1234423-95-0); MG132 (MACKLIN, cat. HY132-59); CHX (MACKLIN, cat. HY-12320, 100 μg/ml for CHX assay); Subasumstat (MACKLIN, TAK-981, cat. HY-111789); Na 3 VO 4 (MACKLIN, cat. 13721-39-6); SU6656 (MACKLIN, cat. HY-B078); K3PO4 (MACKLIN, cat. NONE8222); Protein A/G Magnetic Beads (Beyotime, cat. P2108); Anlotinib (cat. AL3818) dihydrochloride (Aladdin, cat. 1360460-82-7); Cobalt Chloride Solution (MACKLIN, cat. 7791-13-1); anti-mouse PD-1 (Bio X Cell, cat. 857122D1); Anti-mouse lgG (Bio X Cell, cat. 84932331); The full-length coding sequences of YY1, HIF-1α and NUSAP1 (or their variants), as well as IDRs of YY1 and HIF-1α were individually subcloned into a modiffed version of a pGEX vector with 6 × His and EGFP or mCherry at the N-terminus. ..

    Plasmid Preparation:

    Article Title: YY1 Enhances the Stability of HIF-1α Protein by Interacting with NUSAP1 in Macrophages within the Prostate Cancer Microenvironment
    Article Snippet: .. Reagents and antibodies used in this study include: YY1 (Abcam, cat. ab109237, 1:1000 for western blot (WB), 1:200 for immunofluorescent (IF) staining); YY1 (H-10) (Santa Cruz, cat. sc-7341, 1:50 for immunoprecipitation (IP), 1:200 for IF); NUSAP1 (Proteintech, cat. 12024-1-AP, 1:2000 for WB, 1:100 for IP, 1:200 for IF); HIF-1α (abcam, cat. ab51608, 1:1000 for WB, 1:500 for IF, 1:100 for IP); Lyn (Proteintech, cat. 18135-1-AP, 1:1000 for WB); and Tubulin (Proteintech, cat. 11224-1-ap, 1/2000 for WB); SUMO 2/3 (Proteintech, cat. 11251-1-ap, 1:1000 for WB); Ubiquitin (Proteintech, cat. 10201-2-ap, 1:1000 for WB); Flag (Cell signaling Technology, cat. 14793S, 1:1000 for WB, 1:200 for IP); EGFP (Proteintech, cat. 66002-1-lg, 1:2000 for WB, 1:200 for IP); HA (Proteintech, cat. 51064-2-ap, 1:2000 for WB, 1:200 for IP); Phospho-Serine (Immunechem, cat. ICP9806, 1:250 for WB); Phospho-Tyrosine (Immunechem, cat. ICP9805, 1:250 for WB); Phospho-YY1 (Affinity, cat. AF3694, 1:1000 for WB); GAPDH (Acton, cat. 10R-G109A, 1:1000 for WB); F4/80 (Aifang biological, cat. SAF002, 1:500 for mIHC); Goat anti-mouse IgG-HRP (Santa Cruz, cat. sc2005,1:5000 for WB); Goat anti-rabbit IgG-HRP (Santa Cruz, cat. sc-2004, 1:5000 for WB); PE-CD8 (BD Bioscience, cat. 553030, 1:50 for flow cytometry (FC)); APC-CD3 (BD Bioscience, cat. 555275, 1:50 for FC); PE-CD274 (Biolegend, cat. 329706, 1:50 for FC); 1,6-hexanediol (Aladdin, cat. H103708); Tenapanor (TargetMol, T7587, cat. 1234423-95-0); MG132 (MACKLIN, cat. HY132-59); CHX (MACKLIN, cat. HY-12320, 100 μg/ml for CHX assay); Subasumstat (MACKLIN, TAK-981, cat. HY-111789); Na 3 VO 4 (MACKLIN, cat. 13721-39-6); SU6656 (MACKLIN, cat. HY-B078); K3PO4 (MACKLIN, cat. NONE8222); Protein A/G Magnetic Beads (Beyotime, cat. P2108); Anlotinib (cat. AL3818) dihydrochloride (Aladdin, cat. 1360460-82-7); Cobalt Chloride Solution (MACKLIN, cat. 7791-13-1); anti-mouse PD-1 (Bio X Cell, cat. 857122D1); Anti-mouse lgG (Bio X Cell, cat. 84932331); The full-length coding sequences of YY1, HIF-1α and NUSAP1 (or their variants), as well as IDRs of YY1 and HIF-1α were individually subcloned into a modiffed version of a pGEX vector with 6 × His and EGFP or mCherry at the N-terminus. ..



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    A Coomassie blue staining of SERPINB3, SENP3 and NPM1 proteins before and after purification from E-coli . B Western blot analysis of SENP3 and NPM1 after immunoprecipitated with Flag antibody using SENP3, NPM1 and Flag-SERPINB3 purified proteins. Experiments were repeated twice. C Cartoon model of the SENP3-SCCA1 complex predicted by AlphaFold3. D Predicted aligned error (PAE) map of the SENP3-SCCA1 complex predicted by AlphaFold3. E Western blot analysis of SENP3 and truncated SENP3 (Δ265-287) after Co-IP using Flag antibody with purified proteins. F Immunofluorescent analysis of HA tag in PC9 cells overexpressing wild-type SENP3 and SENP3 Δ265-287. Scale bars: 50 μm. G Western blot analysis of NPM1 sumoylation after immunoprecipitated with NPM1 antibody in PC9 cells overexpressing wild-type SENP3 and SENP3 Δ265-287. H Western blot analysis of NPM1 sumoylation after immunocipitated with NPM1 antibody in PC9 cells transfected with Flag-SERPINB3, HA-SENP3 and HA-SENP3(△265-287) viruses as indicated. Experiments were repeated twice. The ratios of <t>SUMO2-NPM1/NPM1</t> were calculated and indicated. I , J Cartoon model of the pentameric NPM1-SENP3 complex predicted by AlphaFold3. K Predicted aligned error (PAE) map of the pentameric NPM1-SENP3 complex predicted by AlphaFold3. L Western blot analysis of SENP3 after Co-IP using Flag antibody with purified proteins. M Lysates from HEK293 cells transfected with plasmids encoding Flag-SERPINB3, HA-NPM1, and His-SENP3 were separated on a Superose 6 Increase 10/300 column. Fractions were resolved by SDS-PAGE and probed with the indicated antibodies. N Lysates from PC9 cells overexpressing Flag-SERPINB3 were separated on a Superose 6 Increase 10/300 column. Fractions were resolved by SDS-PAGE and probed with the indicated antibodies. O Proposed model of SERPINB3/SENP3/NPM1 complex.
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    Image Search Results


    A Coomassie blue staining of SERPINB3, SENP3 and NPM1 proteins before and after purification from E-coli . B Western blot analysis of SENP3 and NPM1 after immunoprecipitated with Flag antibody using SENP3, NPM1 and Flag-SERPINB3 purified proteins. Experiments were repeated twice. C Cartoon model of the SENP3-SCCA1 complex predicted by AlphaFold3. D Predicted aligned error (PAE) map of the SENP3-SCCA1 complex predicted by AlphaFold3. E Western blot analysis of SENP3 and truncated SENP3 (Δ265-287) after Co-IP using Flag antibody with purified proteins. F Immunofluorescent analysis of HA tag in PC9 cells overexpressing wild-type SENP3 and SENP3 Δ265-287. Scale bars: 50 μm. G Western blot analysis of NPM1 sumoylation after immunoprecipitated with NPM1 antibody in PC9 cells overexpressing wild-type SENP3 and SENP3 Δ265-287. H Western blot analysis of NPM1 sumoylation after immunocipitated with NPM1 antibody in PC9 cells transfected with Flag-SERPINB3, HA-SENP3 and HA-SENP3(△265-287) viruses as indicated. Experiments were repeated twice. The ratios of SUMO2-NPM1/NPM1 were calculated and indicated. I , J Cartoon model of the pentameric NPM1-SENP3 complex predicted by AlphaFold3. K Predicted aligned error (PAE) map of the pentameric NPM1-SENP3 complex predicted by AlphaFold3. L Western blot analysis of SENP3 after Co-IP using Flag antibody with purified proteins. M Lysates from HEK293 cells transfected with plasmids encoding Flag-SERPINB3, HA-NPM1, and His-SENP3 were separated on a Superose 6 Increase 10/300 column. Fractions were resolved by SDS-PAGE and probed with the indicated antibodies. N Lysates from PC9 cells overexpressing Flag-SERPINB3 were separated on a Superose 6 Increase 10/300 column. Fractions were resolved by SDS-PAGE and probed with the indicated antibodies. O Proposed model of SERPINB3/SENP3/NPM1 complex.

    Journal: Cell Death & Disease

    Article Title: SERPINB3 enhances NPM1 sumoylation via inhibiting SENP3’s activity and promotes lung tumorigenesis

    doi: 10.1038/s41419-025-08347-9

    Figure Lengend Snippet: A Coomassie blue staining of SERPINB3, SENP3 and NPM1 proteins before and after purification from E-coli . B Western blot analysis of SENP3 and NPM1 after immunoprecipitated with Flag antibody using SENP3, NPM1 and Flag-SERPINB3 purified proteins. Experiments were repeated twice. C Cartoon model of the SENP3-SCCA1 complex predicted by AlphaFold3. D Predicted aligned error (PAE) map of the SENP3-SCCA1 complex predicted by AlphaFold3. E Western blot analysis of SENP3 and truncated SENP3 (Δ265-287) after Co-IP using Flag antibody with purified proteins. F Immunofluorescent analysis of HA tag in PC9 cells overexpressing wild-type SENP3 and SENP3 Δ265-287. Scale bars: 50 μm. G Western blot analysis of NPM1 sumoylation after immunoprecipitated with NPM1 antibody in PC9 cells overexpressing wild-type SENP3 and SENP3 Δ265-287. H Western blot analysis of NPM1 sumoylation after immunocipitated with NPM1 antibody in PC9 cells transfected with Flag-SERPINB3, HA-SENP3 and HA-SENP3(△265-287) viruses as indicated. Experiments were repeated twice. The ratios of SUMO2-NPM1/NPM1 were calculated and indicated. I , J Cartoon model of the pentameric NPM1-SENP3 complex predicted by AlphaFold3. K Predicted aligned error (PAE) map of the pentameric NPM1-SENP3 complex predicted by AlphaFold3. L Western blot analysis of SENP3 after Co-IP using Flag antibody with purified proteins. M Lysates from HEK293 cells transfected with plasmids encoding Flag-SERPINB3, HA-NPM1, and His-SENP3 were separated on a Superose 6 Increase 10/300 column. Fractions were resolved by SDS-PAGE and probed with the indicated antibodies. N Lysates from PC9 cells overexpressing Flag-SERPINB3 were separated on a Superose 6 Increase 10/300 column. Fractions were resolved by SDS-PAGE and probed with the indicated antibodies. O Proposed model of SERPINB3/SENP3/NPM1 complex.

    Article Snippet: SUMO2 antibody for WB (1:1,000, Cell Signaling Technology, #4971S).

    Techniques: Staining, Purification, Western Blot, Immunoprecipitation, Co-Immunoprecipitation Assay, Transfection, SDS Page

    A Western blot analysis of indicated proteins from control and SERPINB3 knockdown PC9 cells. β-actin serves as loading control. Experiments were repeated twice. The ratio of SUMO2/β-actin was calculated and indicated. B Bar graph shows SENP3 activity in control and SERPINB3 knockdown PC9 cells after incubating with hSUMO2-AMC for 50 min. RLU: relative unit. Values are mean ± SD from three independent experiments, * p < 0.05; ** p < 0.01. Student’s unpaired t -test was utilized to determine the statistical significance. Western blot analysis of NPM1 sumoylation after immunoprecipitated with NPM1 antibody in PC9 ( C ) and H1975 ( D ) control, Flag-SERPINB3 overexpression, Flag-SERPINB3 ∆6 overexpression and Flag-Arf overexpression cells. Experiments for each cell line were repeated twice. The ratios of SUMO2-NPM1/NPM1 were calculated and indicated. Western blot analysis of NPM1 sumoylation after immunoprecipitated with NPM1 antibody in PC9 ( E ) and H1975 ( F ) cells transfected with siRNAs targeting SERPINB3, SENP3 and both, as indicated. Transfection of a scrambled oligonucleotide served as a control. Experiments for each cell line were repeated twice. The ratios of SUMO2-NPM1/NPM1 were calculated and indicated.

    Journal: Cell Death & Disease

    Article Title: SERPINB3 enhances NPM1 sumoylation via inhibiting SENP3’s activity and promotes lung tumorigenesis

    doi: 10.1038/s41419-025-08347-9

    Figure Lengend Snippet: A Western blot analysis of indicated proteins from control and SERPINB3 knockdown PC9 cells. β-actin serves as loading control. Experiments were repeated twice. The ratio of SUMO2/β-actin was calculated and indicated. B Bar graph shows SENP3 activity in control and SERPINB3 knockdown PC9 cells after incubating with hSUMO2-AMC for 50 min. RLU: relative unit. Values are mean ± SD from three independent experiments, * p < 0.05; ** p < 0.01. Student’s unpaired t -test was utilized to determine the statistical significance. Western blot analysis of NPM1 sumoylation after immunoprecipitated with NPM1 antibody in PC9 ( C ) and H1975 ( D ) control, Flag-SERPINB3 overexpression, Flag-SERPINB3 ∆6 overexpression and Flag-Arf overexpression cells. Experiments for each cell line were repeated twice. The ratios of SUMO2-NPM1/NPM1 were calculated and indicated. Western blot analysis of NPM1 sumoylation after immunoprecipitated with NPM1 antibody in PC9 ( E ) and H1975 ( F ) cells transfected with siRNAs targeting SERPINB3, SENP3 and both, as indicated. Transfection of a scrambled oligonucleotide served as a control. Experiments for each cell line were repeated twice. The ratios of SUMO2-NPM1/NPM1 were calculated and indicated.

    Article Snippet: SUMO2 antibody for WB (1:1,000, Cell Signaling Technology, #4971S).

    Techniques: Western Blot, Control, Knockdown, Activity Assay, Immunoprecipitation, Over Expression, Transfection

    A Western blot analysis of SERPINB3 expression in Beas2B cells. B Growth curve of Beas2B control and SERPINB3 overexpression cells. Seed 5 × 10 4 Beas2B cells and count for 3 days. Values are mean ± SD from three independent experiments, * p < 0.05, ** p < 0.01. Two way ANOVA test was utilized for multiple group comparison. C Western blot analysis of SERPINB3 expression in PC9 cells. D Growth curve of Beas2B control and SERPINB3 overexpression cells. Seed 1.5 × 10 5 PC9 cells and count for 3 days. Values are mean ± SD from three independent experiments, ** p < 0.01, **** p < 0.0001. Two way ANOVA test was utilized for multiple group comparison. E Xenograft tumors growing from indicated PC9 cell lines were isolated from nude mice after inoculation for 40 days. F Xenograft tumor volume curve of PC9 cells overexpressing empty vector (EV), SERPINB3 and △6. Values are mean ± SEM, N = 5 mice for each group, **** p < 0.0001. Two way ANOVA test was utilized for multiple group comparison. G Western blot analysis of NPM1 sumoylation after immunoprecipitated with NPM1 antibody from xenograft tumor lysates. The ratio of SUMO2-NPM1/NPM1 was indicated.

    Journal: Cell Death & Disease

    Article Title: SERPINB3 enhances NPM1 sumoylation via inhibiting SENP3’s activity and promotes lung tumorigenesis

    doi: 10.1038/s41419-025-08347-9

    Figure Lengend Snippet: A Western blot analysis of SERPINB3 expression in Beas2B cells. B Growth curve of Beas2B control and SERPINB3 overexpression cells. Seed 5 × 10 4 Beas2B cells and count for 3 days. Values are mean ± SD from three independent experiments, * p < 0.05, ** p < 0.01. Two way ANOVA test was utilized for multiple group comparison. C Western blot analysis of SERPINB3 expression in PC9 cells. D Growth curve of Beas2B control and SERPINB3 overexpression cells. Seed 1.5 × 10 5 PC9 cells and count for 3 days. Values are mean ± SD from three independent experiments, ** p < 0.01, **** p < 0.0001. Two way ANOVA test was utilized for multiple group comparison. E Xenograft tumors growing from indicated PC9 cell lines were isolated from nude mice after inoculation for 40 days. F Xenograft tumor volume curve of PC9 cells overexpressing empty vector (EV), SERPINB3 and △6. Values are mean ± SEM, N = 5 mice for each group, **** p < 0.0001. Two way ANOVA test was utilized for multiple group comparison. G Western blot analysis of NPM1 sumoylation after immunoprecipitated with NPM1 antibody from xenograft tumor lysates. The ratio of SUMO2-NPM1/NPM1 was indicated.

    Article Snippet: SUMO2 antibody for WB (1:1,000, Cell Signaling Technology, #4971S).

    Techniques: Western Blot, Expressing, Control, Over Expression, Comparison, Isolation, Plasmid Preparation, Immunoprecipitation

    A Western blot analysis of NPM1 and NPM1 mutants (K230R and K263R) sumoylation in PC9 cells. The ratio of SUMO2-NPM1/tubulin was indicated. B Western blot analysis of NPM1 and NPM1 mutant (K230R and K263R) expression in Beas2B cells. C Growth curve of Beas2B control, NPM1 and NPM1 mutants (K230R and K263R) overexpression cells. Seed 5×10 4 Beas2B cells and count for 3 days. Values are mean ± SD from three independent experiments, *** p < 0.001. Two way ANOVA test was utilized for multiple group comparison. D Western blot analysis of NPM1 and NPM1 mutants (K230R and K263R) expression in PC9 cells. E Growth curve of PC9 control, NPM1 and NPM1 mutants (K230R and K263R) overexpression cells. Seed 1.5×10 5 PC9 cells and count for 3 days. Values are mean ± SD from three independent experiments, **** p < 0.0001. Two way ANOVA test was utilized for multiple group comparison. F Xenograft tumors growing from indicated PC9 cell lines were isolated from nude mice after inoculation for 36 days. G Xenograft tumor volume curve of PC9 control, NPM1 and NPM1 mutants (K230R and K263R) overexpression cells. Values are mean ± SEM, N = 5 mice for each group, ** p < 0.01, *** p < 0.001, **** p < 0.0001. Two way ANOVA test was utilized for multiple group comparison.

    Journal: Cell Death & Disease

    Article Title: SERPINB3 enhances NPM1 sumoylation via inhibiting SENP3’s activity and promotes lung tumorigenesis

    doi: 10.1038/s41419-025-08347-9

    Figure Lengend Snippet: A Western blot analysis of NPM1 and NPM1 mutants (K230R and K263R) sumoylation in PC9 cells. The ratio of SUMO2-NPM1/tubulin was indicated. B Western blot analysis of NPM1 and NPM1 mutant (K230R and K263R) expression in Beas2B cells. C Growth curve of Beas2B control, NPM1 and NPM1 mutants (K230R and K263R) overexpression cells. Seed 5×10 4 Beas2B cells and count for 3 days. Values are mean ± SD from three independent experiments, *** p < 0.001. Two way ANOVA test was utilized for multiple group comparison. D Western blot analysis of NPM1 and NPM1 mutants (K230R and K263R) expression in PC9 cells. E Growth curve of PC9 control, NPM1 and NPM1 mutants (K230R and K263R) overexpression cells. Seed 1.5×10 5 PC9 cells and count for 3 days. Values are mean ± SD from three independent experiments, **** p < 0.0001. Two way ANOVA test was utilized for multiple group comparison. F Xenograft tumors growing from indicated PC9 cell lines were isolated from nude mice after inoculation for 36 days. G Xenograft tumor volume curve of PC9 control, NPM1 and NPM1 mutants (K230R and K263R) overexpression cells. Values are mean ± SEM, N = 5 mice for each group, ** p < 0.01, *** p < 0.001, **** p < 0.0001. Two way ANOVA test was utilized for multiple group comparison.

    Article Snippet: SUMO2 antibody for WB (1:1,000, Cell Signaling Technology, #4971S).

    Techniques: Western Blot, Mutagenesis, Expressing, Control, Over Expression, Comparison, Isolation