Review

α ubc9 (Proteintech)

Proteintech is a verified supplier

About

News

Press Release

Team

Advisors

Partners

Contact

Bioz Stars

Bioz vStars

Buy from Supplier

Structured Review

Proteintech α ubc9

FOXL2 SUMOylation intricately regulates increased expression in CAFs. ( A ) SUMOylation sites of FOXL2 was analyzed by SUMOplot™ analysis. Shown are the top 7 predicted lysine residues; ( B ) In vitro sumoylation assay was employed to confirm the predicted SUMOylation sites using HA-tagged wild-type (WT), or the K25R, K87R, K114R, K150R, and 4KR (where K25, K87, K114 and K150 were all mutated to R). Shown is a representative blot and densitometry analysis of SUMOylated-FOXL2/Total FOXL2; ( C ) The association between SUMOylation and FOXL2 stability was determined by western blotting in HEK-293T cells using α-HA. Cells were transfected with either HA-FOXL2-WT or HA-FOXL2-K25/87R (double mutant, 2KR). The membrane was stripped and re-probed with GAPDH to confirm equivalent loading. Shown is a representative blot and densitometry analysis of three technical replicates; ( D ) Cells were transfected with HA-FOXL2-WT or, HA-FOXL2-2KR ± His-SUMO1. CHX (100 µg/ml) was added to inhibit translation allowing tracking of FOXL2 stability in the presence and absence of His-SUMO1. Blots were probed with α-HA and re-probed with GAPDH to confirm equivalent loading. Shown is a representative blot and densitometry analysis of three technical replicates; ( E ) DMSO (control), MG132 (proteasome inhibitor) or chloroquine (lysosome inhibitor) were added into HEK-293T cells transfected with HA-FOXL2-WT or HA-FOXL2-2KR, before CHX (100 µg/ml) was added. Blots were probed with α-HA and re-probed with GAPDH to confirm equivalent loading. Shown is a representative blot; ( F ) IP assay was used to test the association between FOXL2 SUMOylation and ubiquitination. Lysates obtained from HEK-293T cells transfected with HA-FOXL2-WT or HA-FOXL2-2KR, along with <t>FLAG-UBC9,</t> were immunoprecipitated using α-HA antibody and then probed with α-FLAG antibody. Shown is a representative blot; *, *** P < 0.05, P < 0.001

α Ubc9, supplied by Proteintech, used in various techniques. Bioz Stars score: 93/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/anti+ubc9/UBC9+Antibody/pmc12853930-67-9-21
Average 93 stars, based on 14 article reviews

α ubc9 - by Bioz Stars, 2026-09

93/100 stars

Images

1) Product Images from "FOXL2 + cancer-associated fibroblasts enhances epithelial ovarian cancer development via TGFβ/Smad signaling"

Article Title: FOXL2 + cancer-associated fibroblasts enhances epithelial ovarian cancer development via TGFβ/Smad signaling

Journal: BMC Cancer

doi: 10.1186/s12885-025-15364-6

Figure Legend Snippet: FOXL2 SUMOylation intricately regulates increased expression in CAFs. ( A ) SUMOylation sites of FOXL2 was analyzed by SUMOplot™ analysis. Shown are the top 7 predicted lysine residues; ( B ) In vitro sumoylation assay was employed to confirm the predicted SUMOylation sites using HA-tagged wild-type (WT), or the K25R, K87R, K114R, K150R, and 4KR (where K25, K87, K114 and K150 were all mutated to R). Shown is a representative blot and densitometry analysis of SUMOylated-FOXL2/Total FOXL2; ( C ) The association between SUMOylation and FOXL2 stability was determined by western blotting in HEK-293T cells using α-HA. Cells were transfected with either HA-FOXL2-WT or HA-FOXL2-K25/87R (double mutant, 2KR). The membrane was stripped and re-probed with GAPDH to confirm equivalent loading. Shown is a representative blot and densitometry analysis of three technical replicates; ( D ) Cells were transfected with HA-FOXL2-WT or, HA-FOXL2-2KR ± His-SUMO1. CHX (100 µg/ml) was added to inhibit translation allowing tracking of FOXL2 stability in the presence and absence of His-SUMO1. Blots were probed with α-HA and re-probed with GAPDH to confirm equivalent loading. Shown is a representative blot and densitometry analysis of three technical replicates; ( E ) DMSO (control), MG132 (proteasome inhibitor) or chloroquine (lysosome inhibitor) were added into HEK-293T cells transfected with HA-FOXL2-WT or HA-FOXL2-2KR, before CHX (100 µg/ml) was added. Blots were probed with α-HA and re-probed with GAPDH to confirm equivalent loading. Shown is a representative blot; ( F ) IP assay was used to test the association between FOXL2 SUMOylation and ubiquitination. Lysates obtained from HEK-293T cells transfected with HA-FOXL2-WT or HA-FOXL2-2KR, along with FLAG-UBC9, were immunoprecipitated using α-HA antibody and then probed with α-FLAG antibody. Shown is a representative blot; *, *** P < 0.05, P < 0.001

Techniques Used: Expressing, In Vitro, Western Blot, Transfection, Mutagenesis, Membrane, Control, Ubiquitin Proteomics, Immunoprecipitation

FOXL2 SUMOylation requires SUMO1 and UBC9/UBE2I. ( A ) Putative interaction of FOXL2 with SUMO1, SUMO2, SUMO3, and SUMO4 was detected by String analysis ( https://cn.string-db.org/ ); ( B ) In vitro sumoylation assay was employed to confirm String analysis’s prediction of SUMO1, HEK-293T cells were transfected with HA-FOXL2-WT, FLAG-UBC9, and His-SUMO1-4. In vitro SUMOylation assay using Ni 2+ -NTA pull-down determined that FOXL2 was mainly modified by SUMO1. Shown is a representative blot; ( C ) In vitro sumoylation assay was employed to determine whether UBC9 is compulsorily required for FOXL2 SUMOylation. HEK-293T cells were transfected with HA-FOXL2-WT and His-SUMO1 ± FLAG-UBC9. In vitro SUMOylation using Ni 2+ -NTA pull-down assay determined that UBC9 is required for FOXL2 SUMOylation. Shown is a representative blot; ( D ) In vitro sumoylation assay was employed to determine whether UBC9 is compulsorily required for FOXL2 SUMOylation in CAFs. CAFs were transduced using either a non-targeting control shRNA or shRNA targeting UBC9 . Transduced cells were transfected with HA-FOXL2-WT and His-SUMO1. In vitro SUMOylation using Ni 2+ -NTA pull-down assay determined that UBC9 is required for FOXL2 SUMOylation. Shown is a representative blot

Figure Legend Snippet: FOXL2 SUMOylation requires SUMO1 and UBC9/UBE2I. ( A ) Putative interaction of FOXL2 with SUMO1, SUMO2, SUMO3, and SUMO4 was detected by String analysis ( https://cn.string-db.org/ ); ( B ) In vitro sumoylation assay was employed to confirm String analysis’s prediction of SUMO1, HEK-293T cells were transfected with HA-FOXL2-WT, FLAG-UBC9, and His-SUMO1-4. In vitro SUMOylation assay using Ni 2+ -NTA pull-down determined that FOXL2 was mainly modified by SUMO1. Shown is a representative blot; ( C ) In vitro sumoylation assay was employed to determine whether UBC9 is compulsorily required for FOXL2 SUMOylation. HEK-293T cells were transfected with HA-FOXL2-WT and His-SUMO1 ± FLAG-UBC9. In vitro SUMOylation using Ni 2+ -NTA pull-down assay determined that UBC9 is required for FOXL2 SUMOylation. Shown is a representative blot; ( D ) In vitro sumoylation assay was employed to determine whether UBC9 is compulsorily required for FOXL2 SUMOylation in CAFs. CAFs were transduced using either a non-targeting control shRNA or shRNA targeting UBC9 . Transduced cells were transfected with HA-FOXL2-WT and His-SUMO1. In vitro SUMOylation using Ni 2+ -NTA pull-down assay determined that UBC9 is required for FOXL2 SUMOylation. Shown is a representative blot

Techniques Used: In Vitro, Transfection, Modification, Pull Down Assay, Control, shRNA

other:

Article Title: Pharmacologic Induction of ERα SUMOylation Disrupts Its Chromatin Binding.
Article Snippet: The following primary antibodies were used, diluted in 5% nonfat milk PBST: anti-ERα (Abcam, ab16660, 1:2500), anti-FLAG-HRP (SigmaAldrich, A8592, 1:10 000), anti-HA (Cell Signaling Technology, 3724S, 1:2500), anti-UBC9 (Proteintech, 10070−1-AP, 1:5000), anti-His (Origene, TA150088, 1:4000), anti-β-ACTIN-HRP (Huaxingbio, HX18271, 1:10 000), and anti-GAPDH-HRP (Abclonal, AC035, 1:8000).

Article Title: The Interaction between SARS-CoV-2 Nucleocapsid Protein and UBC9 Inhibits MAVS Ubiquitination by Enhancing Its SUMOylation
Article Snippet: Anti-GFP (66002-1-Ig), anti-GST (66001-2-Ig), anti-UBC9 (60201-1-Ig) and anti-GAPDH (60004-1-Ig) antibodies were purchased from Proteintech Group (Wuhan, China).

Article Title: Pharmacologic Induction of ERα SUMOylation Disrupts Its Chromatin Binding
Article Snippet: The following primary antibodies were used, diluted in 5% nonfat milk PBST: anti-ERα (Abcam, ab16660, 1:2500), anti-FLAG-HRP (Sigma-Aldrich, A8592, 1:10 000), anti-HA (Cell Signaling Technology, 3724S, 1:2500), anti-UBC9 (Proteintech, 10070–1-AP, 1:5000), anti-His (Origene, TA150088, 1:4000), anti-β-ACTIN-HRP (Huaxingbio, HX18271, 1:10 000), and anti-GAPDH-HRP (Abclonal, AC035, 1:8000).

Article Title: miR-122-3p targets UBE2I to regulate the immunosuppression of liver cancer and the intervention of Liujunzi formula.
Article Snippet: Ethnopharmacological relevance: Liujunzi formula has been used to treat liver cancer in China for many years, but its underlying mechanism remains unclear.. We previously found that decreased expression of miR-122-3p was associated with liver cancer.. In this study, we aimed to explore the target of miR-122-3p and the effect of the Liujunzi formula on miR-122-3p and its downstream events in liver cancer.

Article Title: Identification of proximal SUMO-dependent interactors using SUMO-ID
Article Snippet: Antibodies used: anti-biotin-HRP (1/1000; Cat#7075S), anti-Myc (1/1000; Cat#2276S), anti-alpha-Actinin (1/5000; Cat#6487S) (Cell Signaling Technology); anti-Flag (1/1000; Cat#F1804), anti-GTF2I (1/1000; Cat#HPA026638) (Sigma-Aldrich); anti-BirA (1/1000; Cat#11582-T16; SinoBiological); Proteintech antibodies: anti-IRF2BP2 (1/1000; Cat#18847-1-AP), anti-UBC9 (1/1000; Cat#14837-1-AP), anti-TRIM24 (1/1000; Cat#14208-1-AP), anti-TRIM33 (1/1000; Cat#55374-1-AP), anti-PIAS2 (1/1000; Cat#16074-1-AP), anti-PIAS4 (1/1000; Cat#14242-1-AP), anti-GATAD2B (1/1000; Cat#25679-1-AP), anti-MTA2 (1/1000; Cat#17554-1-AP), anti-RBBP4 (1/1000; Cat#20364-1-AP), anti-PML (1/1000; Cat#21041-1-AP), anti-GAPDH (1/5000; Cat#60004-1-Ig), anti-beta-Actin (1/5000; Cat#66009-1-Ig), anti-alpha-Tubulin (1/5000; Cat#66031-1-Ig); anti-PML (1/1000; Cat#A301-167A) (Bethyl); anti-GFP (1/1000; Cat#sc-8334) (SantaCruz); anti-Mouse-HRP (1/5000; Cat#115-035-062), anti-Rabbit-HRP (1/5000; Cat#111-035-045) (Jackson ImmunoResearch).

Article Title: The Interaction between SARS-CoV-2 Nucleocapsid Protein and UBC9 Inhibits MAVS Ubiquitination by Enhancing Its SUMOylation.
Article Snippet: Anti-GFP (66002-1-Ig), anti-GST (66001-2-Ig), anti-UBC9 (60201-1-Ig) and anti-GAPDH (60004-1-Ig) antibodies were purchased from Proteintech Group (Wuhan, China).

Article Title: A novel UVA-associated circUBE2I mediates ferroptosis in HaCaT cells.
Article Snippet: 1Department of Anesthesiology, The Affiliated Hospital, Southwest Medical University, Luzhou, Sichuan Province, China 2Anesthesiology and Critical Care Medicine Key Laboratory of Luzhou, The Affiliated Hospital, Southwest Medical University, Luzhou, Sichuan Province, China 3Department of Dermatology, The Affiliated Hospital, Southwest Medical University, Luzhou, Sichuan Province, China 4Department of Dermatology, Suining First People's Hospital, Suining, Sichuan, China 5Moutai Institute, Renhuai, Guizhou Province, China

Incubation:

Article Title: Antimony exposure induces bladder cancer metastasis via activation of the SUMOylation-dependent TGF-β/Smad2/3 axis.
Article Snippet: Antimony is an important industrial raw material, known as the "industrial monosodium glutamate", and is widely used in alloys, electronics, chemical engineering, and other fields.. Due to its extensive applications, its environmental toxicity and carcinogenic risk have attracted increasing attention.. In recent years, numerous studies have reported the carcinogenic risk of antimony, but its role in bladder cancer and the underlying molecular mechanisms have not been fully elucidated.

Image Search Results

a Immunofluorescence images and quantification (n = 20) of SUMO1 (green) and SUMO2/3 (red) conjugation in RAW264.7 cells with or without STM 14028S infection (4 hpi). Scale bar, 10 µm. b Core SUMO cycle enzymes and their corresponding primary genes (blue). c, d Transcriptomic and proteomic analyses of SUMO cycle enzyme expression in RAW264.7 cells infected with STM 14028S vs uninfected controls (4 hpi; n = 3). e UBC9 mRNA expression in RAW264.7 cells during STM 14028S infection (0-6 hpi; n = 3). f Immunoblot analysis and quantification of UBC9 protein levels in STM 14028S-infected RAW264.7 cells (0-6 hpi; n = 3). *P < 0.05; ***P < 0.001; ns, not significant.

Journal: bioRxiv

Article Title: A bacterial effector blocks SUMOylation by steric occlusion of UBC9 via arginine-GlcNAcylation

doi: 10.64898/2026.03.06.710069

Figure Lengend Snippet: a Immunofluorescence images and quantification (n = 20) of SUMO1 (green) and SUMO2/3 (red) conjugation in RAW264.7 cells with or without STM 14028S infection (4 hpi). Scale bar, 10 µm. b Core SUMO cycle enzymes and their corresponding primary genes (blue). c, d Transcriptomic and proteomic analyses of SUMO cycle enzyme expression in RAW264.7 cells infected with STM 14028S vs uninfected controls (4 hpi; n = 3). e UBC9 mRNA expression in RAW264.7 cells during STM 14028S infection (0-6 hpi; n = 3). f Immunoblot analysis and quantification of UBC9 protein levels in STM 14028S-infected RAW264.7 cells (0-6 hpi; n = 3). *P < 0.05; ***P < 0.001; ns, not significant.

Article Snippet: Unique primary antibodies used in this study included UBC9 (CST, #4786), SUMO1 (Proteintech, 67557-1-lg), SUMO2/3 (Proteintech, 67154-1-lg), arginine-GlcNAcylation antibody (Abcam, EPR18251), Myd88 (Proteintech, 67969-1-lg), HSPA8 (Proteintech, 10654-1-AP), PDCD4 (Proteintech, 84162-3-RR).

Techniques: Immunofluorescence, Conjugation Assay, Infection, Expressing, Western Blot

a Schematic of the Y2H screening workflow and pairwise interaction validation strategy. b Representative positive colonies on selective plates; four colonies (blue arrows) correspond to Ube2i (encoding UBC9). c Sequencing analysis of positive clones showing that four independent clones encode Ube2i (orange). d Pairwise interaction assay validating the interaction between SseK1 and UBC9. e Co-IP in HEK293T cells confirming the interaction between SseK1 and UBC9. f In vitro pull-down assay demonstrating direct binding between purified SseK1 and UBC9. g BLI analysis showing the binding kinetics between SseK1 and UBC9 in vitro .

Journal: bioRxiv

Article Title: A bacterial effector blocks SUMOylation by steric occlusion of UBC9 via arginine-GlcNAcylation

doi: 10.64898/2026.03.06.710069

Figure Lengend Snippet: a Schematic of the Y2H screening workflow and pairwise interaction validation strategy. b Representative positive colonies on selective plates; four colonies (blue arrows) correspond to Ube2i (encoding UBC9). c Sequencing analysis of positive clones showing that four independent clones encode Ube2i (orange). d Pairwise interaction assay validating the interaction between SseK1 and UBC9. e Co-IP in HEK293T cells confirming the interaction between SseK1 and UBC9. f In vitro pull-down assay demonstrating direct binding between purified SseK1 and UBC9. g BLI analysis showing the binding kinetics between SseK1 and UBC9 in vitro .

Techniques: Biomarker Discovery, Sequencing, Clone Assay, Co-Immunoprecipitation Assay, In Vitro, Pull Down Assay, Binding Assay, Purification

a HEK293T cells expressing Flag-UBC9 were infected with STM WT or Δ sseK1 , followed by immunoprecipitation and immunoblot analysis of UBC9 Arg-GlcNAcylation. b In vitro enzymatic assays showing Arg-GlcNAcylation of bacterially expressed 6×His-UBC9, detected by immunoblotting. c MS/MS spectrum of Arg-GlcNAcylated Flag-UBC9 peptides. Corresponding b and y ions are indicated along the peptide sequence above the spectrum. d Immunoblot analysis and quantification (normalized to Coomassie-stained protein, n = 3) of Arg-GlcNAcylation of UBC9 and UBC9 R17A. **P < 0.01.

Journal: bioRxiv

Article Title: A bacterial effector blocks SUMOylation by steric occlusion of UBC9 via arginine-GlcNAcylation

doi: 10.64898/2026.03.06.710069

Figure Lengend Snippet: a HEK293T cells expressing Flag-UBC9 were infected with STM WT or Δ sseK1 , followed by immunoprecipitation and immunoblot analysis of UBC9 Arg-GlcNAcylation. b In vitro enzymatic assays showing Arg-GlcNAcylation of bacterially expressed 6×His-UBC9, detected by immunoblotting. c MS/MS spectrum of Arg-GlcNAcylated Flag-UBC9 peptides. Corresponding b and y ions are indicated along the peptide sequence above the spectrum. d Immunoblot analysis and quantification (normalized to Coomassie-stained protein, n = 3) of Arg-GlcNAcylation of UBC9 and UBC9 R17A. **P < 0.01.

Techniques: Expressing, Infection, Immunoprecipitation, Western Blot, In Vitro, Tandem Mass Spectroscopy, Sequencing, Staining

a Crystal structure of the UBC9-SUMO1 complex (PDB: 2UYZ). b Predicted structure of the UBC9-SUMO2 complex generated using AlphaFold3. Enlarged views of the UBC9-SUMO1 and UBC9-SUMO2 interfaces are highlighted in black boxes. Residues forming hydrogen bonds with UBC9 R17 are shown in stick representation. c Purification of 6×His-UBC9 from E. coli BL21(DE3) co-expressing pSseK1 and pUBC9, followed by immunoblot analysis of UBC9 Arg-GlcNAcylation. d Purification of SUMO1 and SUMO2 proteins from E. coli BL21(DE3) harboring pSUMO1 or pSUMO2, respectively. M, molecular weight marker. e, f MST analysis of the binding affinities between SUMO1 or SUMO2 and unmodified UBC9 or Arg-GlcNAcylated UBC9.

Journal: bioRxiv

Article Title: A bacterial effector blocks SUMOylation by steric occlusion of UBC9 via arginine-GlcNAcylation

doi: 10.64898/2026.03.06.710069

Figure Lengend Snippet: a Crystal structure of the UBC9-SUMO1 complex (PDB: 2UYZ). b Predicted structure of the UBC9-SUMO2 complex generated using AlphaFold3. Enlarged views of the UBC9-SUMO1 and UBC9-SUMO2 interfaces are highlighted in black boxes. Residues forming hydrogen bonds with UBC9 R17 are shown in stick representation. c Purification of 6×His-UBC9 from E. coli BL21(DE3) co-expressing pSseK1 and pUBC9, followed by immunoblot analysis of UBC9 Arg-GlcNAcylation. d Purification of SUMO1 and SUMO2 proteins from E. coli BL21(DE3) harboring pSUMO1 or pSUMO2, respectively. M, molecular weight marker. e, f MST analysis of the binding affinities between SUMO1 or SUMO2 and unmodified UBC9 or Arg-GlcNAcylated UBC9.

Techniques: Generated, Purification, Expressing, Western Blot, Molecular Weight, Marker, Binding Assay

a Immunoblot analysis of UBC9 Arg-GlcNAcylation mediated by SseK1, SseK2, or SseK3. b AlphaFold-predicted structures of SseK1 (aa 29-336, red), SseK2 (aa 29-348, yellow), and SseK3 (aa 29-335, green), shown with structural alignment using PyMOL. c AlphaFold-modeled structure of the SseK1-UBC9 complex. The enlarged view of the lid-domain region is boxed in black. Residues in the SseK1 lid domain forming hydrogen bonds with UBC9 are shown in stick representation. d Sequence alignment of SseK1, SseK2, and SseK3 generated using ESPript 3.0. e Immunoblot analysis of UBC9 Arg-GlcNAcylation mediated by SseK1, SseK1 A332_Q336del, SseK3, and SseK3 R332delinsARHVQ. f Immunoblot analysis of SUMO2/3 conjugation in RAW264.7 cells infected with STM Δ sseK1 , STM Δ sseK1 complemented with SseK1, or STM Δ sseK1 complemented with either SseK1 D223_D225delinsAAA or SseK1 A332_Q336del (4 hpi). g Phylogenetic analysis of Salmonella Typhimurium SseK1 homologs. Protein sequences homologous to SseK1 (UniProt accession: A0A0H3NK84) were identified using BLASTP analysis against the UniProtKB reference proteomes and Swiss-Prot databases. Sequences with an E-value < 0.05 were selected for phylogenetic analysis. The phylogenetic tree was constructed and visualized using the Interactive Tree of Life (iTOL) online tool.

Journal: bioRxiv

Article Title: A bacterial effector blocks SUMOylation by steric occlusion of UBC9 via arginine-GlcNAcylation

doi: 10.64898/2026.03.06.710069

Figure Lengend Snippet: a Immunoblot analysis of UBC9 Arg-GlcNAcylation mediated by SseK1, SseK2, or SseK3. b AlphaFold-predicted structures of SseK1 (aa 29-336, red), SseK2 (aa 29-348, yellow), and SseK3 (aa 29-335, green), shown with structural alignment using PyMOL. c AlphaFold-modeled structure of the SseK1-UBC9 complex. The enlarged view of the lid-domain region is boxed in black. Residues in the SseK1 lid domain forming hydrogen bonds with UBC9 are shown in stick representation. d Sequence alignment of SseK1, SseK2, and SseK3 generated using ESPript 3.0. e Immunoblot analysis of UBC9 Arg-GlcNAcylation mediated by SseK1, SseK1 A332_Q336del, SseK3, and SseK3 R332delinsARHVQ. f Immunoblot analysis of SUMO2/3 conjugation in RAW264.7 cells infected with STM Δ sseK1 , STM Δ sseK1 complemented with SseK1, or STM Δ sseK1 complemented with either SseK1 D223_D225delinsAAA or SseK1 A332_Q336del (4 hpi). g Phylogenetic analysis of Salmonella Typhimurium SseK1 homologs. Protein sequences homologous to SseK1 (UniProt accession: A0A0H3NK84) were identified using BLASTP analysis against the UniProtKB reference proteomes and Swiss-Prot databases. Sequences with an E-value < 0.05 were selected for phylogenetic analysis. The phylogenetic tree was constructed and visualized using the Interactive Tree of Life (iTOL) online tool.

Techniques: Western Blot, Sequencing, Generated, Conjugation Assay, Infection, Construct

Approximately two hours after entering macrophages, Salmonella activates its T3SS-2 to deliver effector proteins, including SseK1, SseK2 and SseK3, into the cytoplasm. Among these effectors, SseK1 uniquely engages UBC9 through its unique lid domain and catalyzes arginine-GlcNAcylation of UBC9 at residue R17. This modification prevents UBC9 from forming the UBC9-SUMO thioester intermediate, thereby blocking SUMO conjugation onto a broad set of antimicrobial substrates and impairing their activity, stability or localization. By blocking this SUMO-dependent arm of the host immune response, Salmonella evades host antimicrobial defenses, thereby promoting its survival and enhancing virulence.

Journal: bioRxiv

Article Title: A bacterial effector blocks SUMOylation by steric occlusion of UBC9 via arginine-GlcNAcylation

doi: 10.64898/2026.03.06.710069

Figure Lengend Snippet: Approximately two hours after entering macrophages, Salmonella activates its T3SS-2 to deliver effector proteins, including SseK1, SseK2 and SseK3, into the cytoplasm. Among these effectors, SseK1 uniquely engages UBC9 through its unique lid domain and catalyzes arginine-GlcNAcylation of UBC9 at residue R17. This modification prevents UBC9 from forming the UBC9-SUMO thioester intermediate, thereby blocking SUMO conjugation onto a broad set of antimicrobial substrates and impairing their activity, stability or localization. By blocking this SUMO-dependent arm of the host immune response, Salmonella evades host antimicrobial defenses, thereby promoting its survival and enhancing virulence.

Techniques: Residue, Modification, Blocking Assay, Conjugation Assay, Activity Assay

Journal: BMC Cancer

Article Title: FOXL2 + cancer-associated fibroblasts enhances epithelial ovarian cancer development via TGFβ/Smad signaling

doi: 10.1186/s12885-025-15364-6

Figure Lengend Snippet: FOXL2 SUMOylation intricately regulates increased expression in CAFs. ( A ) SUMOylation sites of FOXL2 was analyzed by SUMOplot™ analysis. Shown are the top 7 predicted lysine residues; ( B ) In vitro sumoylation assay was employed to confirm the predicted SUMOylation sites using HA-tagged wild-type (WT), or the K25R, K87R, K114R, K150R, and 4KR (where K25, K87, K114 and K150 were all mutated to R). Shown is a representative blot and densitometry analysis of SUMOylated-FOXL2/Total FOXL2; ( C ) The association between SUMOylation and FOXL2 stability was determined by western blotting in HEK-293T cells using α-HA. Cells were transfected with either HA-FOXL2-WT or HA-FOXL2-K25/87R (double mutant, 2KR). The membrane was stripped and re-probed with GAPDH to confirm equivalent loading. Shown is a representative blot and densitometry analysis of three technical replicates; ( D ) Cells were transfected with HA-FOXL2-WT or, HA-FOXL2-2KR ± His-SUMO1. CHX (100 µg/ml) was added to inhibit translation allowing tracking of FOXL2 stability in the presence and absence of His-SUMO1. Blots were probed with α-HA and re-probed with GAPDH to confirm equivalent loading. Shown is a representative blot and densitometry analysis of three technical replicates; ( E ) DMSO (control), MG132 (proteasome inhibitor) or chloroquine (lysosome inhibitor) were added into HEK-293T cells transfected with HA-FOXL2-WT or HA-FOXL2-2KR, before CHX (100 µg/ml) was added. Blots were probed with α-HA and re-probed with GAPDH to confirm equivalent loading. Shown is a representative blot; ( F ) IP assay was used to test the association between FOXL2 SUMOylation and ubiquitination. Lysates obtained from HEK-293T cells transfected with HA-FOXL2-WT or HA-FOXL2-2KR, along with FLAG-UBC9, were immunoprecipitated using α-HA antibody and then probed with α-FLAG antibody. Shown is a representative blot; *, *** P < 0.05, P < 0.001

Article Snippet: The different primary antibodies used were α-SUMO1 (67559-1-Ig, 1:3000), α-UBC9 (10070-1-AP, 1:2000), α-Vimentin (10366-1-AP, 1:5000), α-N-cadherin (22018-1-AP, 1:5000), α-E-cadherin (20874-1-AP, 1:20000) (Proteintech, Wuhan, China); α-HA (A02041, 1:800), α-His (A02050, 1:800), α-Flag (A02010, 1:800) (Abbkine, Wuhan, China); α-FOXL2 (ab246511, 1:1000), α-Snail (ab216347, 1:1000) (Abcam, Cambridge, MA, USA); and, α-Smad2/3 (D7G7, 1:1000), α-p-Smad2 (Ser465/467)/Smad3 (Ser423/425) (D27F4, 1:1000) (CST, Boston, MA, USA).

Techniques: Expressing, In Vitro, Western Blot, Transfection, Mutagenesis, Membrane, Control, Ubiquitin Proteomics, Immunoprecipitation

Journal: BMC Cancer

Article Title: FOXL2 + cancer-associated fibroblasts enhances epithelial ovarian cancer development via TGFβ/Smad signaling

doi: 10.1186/s12885-025-15364-6

Figure Lengend Snippet: FOXL2 SUMOylation requires SUMO1 and UBC9/UBE2I. ( A ) Putative interaction of FOXL2 with SUMO1, SUMO2, SUMO3, and SUMO4 was detected by String analysis ( https://cn.string-db.org/ ); ( B ) In vitro sumoylation assay was employed to confirm String analysis’s prediction of SUMO1, HEK-293T cells were transfected with HA-FOXL2-WT, FLAG-UBC9, and His-SUMO1-4. In vitro SUMOylation assay using Ni 2+ -NTA pull-down determined that FOXL2 was mainly modified by SUMO1. Shown is a representative blot; ( C ) In vitro sumoylation assay was employed to determine whether UBC9 is compulsorily required for FOXL2 SUMOylation. HEK-293T cells were transfected with HA-FOXL2-WT and His-SUMO1 ± FLAG-UBC9. In vitro SUMOylation using Ni 2+ -NTA pull-down assay determined that UBC9 is required for FOXL2 SUMOylation. Shown is a representative blot; ( D ) In vitro sumoylation assay was employed to determine whether UBC9 is compulsorily required for FOXL2 SUMOylation in CAFs. CAFs were transduced using either a non-targeting control shRNA or shRNA targeting UBC9 . Transduced cells were transfected with HA-FOXL2-WT and His-SUMO1. In vitro SUMOylation using Ni 2+ -NTA pull-down assay determined that UBC9 is required for FOXL2 SUMOylation. Shown is a representative blot

Techniques: In Vitro, Transfection, Modification, Pull Down Assay, Control, shRNA

Review

α ubc9 (Proteintech)

Structured Review

Images

1) Product Images from "FOXL2 + cancer-associated fibroblasts enhances epithelial ovarian cancer development via TGFβ/Smad signaling"

Related Articles

other:

Incubation:

Similar Products

Image Search Results