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ABclonal Biotechnology resource source identifier antibodies rabbit monoclonal anti sumo2
Resource Source Identifier Antibodies Rabbit Monoclonal Anti Sumo2, supplied by ABclonal Biotechnology, used in various techniques. Bioz Stars score: 94/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/sumo2/SUMO2+Rabbit+pAb/pm42603277-318-2-9
Average 94 stars, based on 7 article reviews

resource source identifier antibodies rabbit monoclonal anti sumo2 - by Bioz Stars, 2026-11

94/100 stars

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other:

Article Title: KSHV hijacks the antiviral kinase IKKε to initiate lytic replication
Article Snippet: The cytotoxicity effects of selected inhibitors were determined by Cell Viability Assay with Cell Counting Kit-8 (Beyotime Biotechnology, #C0037).

Article Title: KSHV hijacks the antiviral kinase IKKε to initiate lytic replication.
Article Snippet: Anti-SUMO1 (#A19121), SUMO2/3 (#A5066), SP100 (#A5851) antibodies were purchased from ABclonal.

Incubation:

Article Title: Alone or in combination, hyaluronic acid and chondroitin sulfate alleviate ECM degradation in osteoarthritis by inhibiting the NF-κB pathway
Article Snippet: After transferring the samples to a PVDF membrane (WJ001S, epizyme, Shanghai China), the membrane was closed with 5% skimmed milk powder, and the samples were incubated with anti-IκBα (A11168, 1:1000, Abclonal, Wuhan, China), p-IκBα (AP0614, 1:1000, Abclonal, Wuhan, China), p65 (A22684, 1:1000, Abclonal, Wuhan, China), and p-65 (AP0123, 1:1000, Abclonal, Wuhan, China). .. Shanghai China), Collagen-II (A24863, 1:1000, Abclonal, Wuhan, China), Aggrecan (MA3-16888, 1:1000, ThermoFisher, Waltham, MA, USA) and GAPDH (AC001, 1:1000, Abclonal, Wuhan, China) primary antibodies were incubated at 4 °C overnight. .. Then it was incubated with HRP-coupled secondary antibody (1:2000, Proteintech) at room temperature for 2 h. It was washed with TBST solution for 5 min and developed using BeyoECL Star (G2020, Servicebio, Wuhan, China) assay for 30 s. The gray values of the bands were analyzed using ImageJ software (V1.8.0.112, NIH, Madison, WI, USA).

Article Title: Zmiz1-Mediated SUMOylation of NLRP3 Inflammasome Regulates Satellite Glial Cell Activation and Neuronal Autophagy in Trigeminal Neuralgia
Article Snippet: .. Subsequently, proteins were transferred to a PVDF membrane using the wet transfer method, blocked with 5% non-fat milk at room temperature for 1 h. The PVDF membrane was then incubated overnight at 4 °C with the following primary antibodies: Flag (AE005, abclonal), HA (AE008, abclonal), NLRP3 (A5652, abclonal), Zmiz1 (ABIN2781101, antibodies-online), IL-1β (A16288, abclonal), cleaved IL-1β (AF4006, affbiotech), caspase-1 (A16792, abclonal), cleaved caspase-1 (AF4005, affbiotech), SUMO1 (A2130, abclonal), SUMO2/3 (A2571, abclonal), GFAP (A19058, abclonal), LC3-I/LC3-II (A5618, abclonal), p62 (A19700, abclonal), and ACTIN (AC026, abclonal). .. The membrane was washed thrice with TBST for 10 min each, then incubated for 1 h with an HRP-conjugated goat anti-rabbit IgG H&L secondary antibody (ab97051, 1:2000, Abcam, Cambridge, UK).

Article Title: SUMOylation and ubiquitination reciprocally regulate SMCHD1 antiviral activity against herpes simplex virus 1
Article Snippet: .. The diluted samples were incubated with anti-FLAG beads or with protein A/G beads together with rabbit control IgG (2 μg), anti-SUMO1 antibody (2 μg; A19121, Abclonal), or anti-SUMO2/3 antibody (2 μg; A22734, Abclonal) overnight at 4°C. ..

Membrane:

Western Blot:

Article Title: Human ANP32A/B are SUMOylated and utilized by avian influenza virus NS2 protein to overcome species-specific restriction
Article Snippet: Cells were harvested for analysis 24 hours after the second transfection step. siRNA oligonucleotides against SENP1 (siRNA1: 5’-GCGCCAGAUUGAAGAACAGAA-3’; siRNA2: 5’-UGACCAUUACACGCAAAGAUA-3’), PIAS2(siRNA1:5’-GCCAUGUUAUUACAGAGAUUA-3’;siRNA2:5’-GCUGCUAUUCCGCCUUCAUUA-3’) and non-targeting control (5’-UUCUCCGAACGUGUCACGU-3’) were purchased from Seven Innovation (China, Beijing) Biotechnology. .. Western blot analysis was conducted following established protocols using the following antibodies: rabbit anti-Flag (Sigma, F7425), mouse anti-Flag (Sigma, F1804), rabbit anti-HA (Sigma, H6908), rabbit anti-ACTB (Abclonal, AC026), mouse anti-ACTB (Abclonal, AC004), rabbit anti-Myc (Abclonal, AE070), rabbit anti-SUMO1 (Abclonal, A19121), rabbit anti-SUMO2/3 (Abclonal, A5066), rabbit anti-SENP1(Abcam, ab108981), mouse anti-His (Proteintech, 66005-1-Ig), rabbit anti-V5 (Proteintech, 14440-1-AP), rabbit anti-SENP1 (Proteintech, 25349-1-AP), rabbit anti-PIAS2 (Proteintech, 16074-1-AP), rabbit anti-GST (Proteintech, 10000-0-AP), rabbit anti-ANP32A (Proteintech,15810-1-AP), rabbit anti-ANP32B (Proteintech, 10843-1-AP), mouse anti-ANP32A (Proteintech, 67687-1-Ig), mouse anti-ANP32B (Proteintech, 66160-1-Ig), rabbit anti-influenza A virus NS2 (GeneTex, GTX125953), rabbit anti-influenza B virus NP (GeneTex, GTX128538), rabbit anti-influenza A virus PB2 (GeneTex, GTX125926), rabbit anti-influenza A virus PA (GeneTex, GTX118991), mouse anti-influenza A virus PA (prepared in our laboratory, 1:5000 for WB), mouse anti-influenza A virus NP (prepared in our laboratory, 1:5000 for WB), Biotin-conjugated Affinipure Goat Anti-Rabbit IgG(H + L) (Proteintech, SA00004-2), DyLight 800-labeled Anti-Mouse IgG (H + L) Antibody (KPL, 5230-0415), DyLight 680-labeled Anti-Rabbit IgG (H + L) Antibody (KPL, 5230-0402) and DyLightTM 680-labeled streptavidin (KPL, 5270-0025). ..

Virus:

Article Title: Dietary soy fiber limits Salmonella infection by disrupting the self-reinforcing loop between pathogen virulence and host inflammation.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Rabbit monoclonal anti-SUMO2/3 ABclonal Cat# A22734; RRID: AB_3740081 Rabbit monoclonal anti-STAT1α ImmunoWay Cat# YM8042; RRID: AB_3740082 Rabbit monoclonal anti-STAT1 (Phospho Tyr701) ImmunoWay Cat# YM8746; RRID: AB_3740083 Rabbit monoclonal anti-c-Fos Cell Signaling Technology Cat# 2250; RRID: AB_2247211 Rabbit monoclonal anti-c-Jun Cell Signaling Technology Cat# 9165; RRID: AB_2130165 Rabbit monoclonal anti-TriMethyl-Histone H3-K4 ABclonal Cat# A22146; RRID: AB_3066542 Rabbit polyclonal anti-beta Actin Abcam Cat# ab8227; RRID: AB_2305186 HRP goat anti-rabbit IgG Abcam Cat# ab7090; RRID: AB_955417 Bacterial and virus strains S. Enteritidis SZ20HS10 Wang et al.41 N/A S. Enteritidis 13076 Laboratory stock N/A ΔttrA S. Enteritidis SZ20HS10 ttrA deletion mutant This study N/A ΔtorC S. Enteritidis SZ20HS10 torC deletion mutant This study N/A Δfur S. Enteritidis SZ20HS10 fur deletion mutant This study N/A ΔhilD S. Enteritidis SZ20HS10 hilD deletion mutant This study N/A Akkermansia muciniphila ATCC BAA-835 mingzhoubio B336076 Biological samples Plasma from mice This study N/A Feces from mice This study N/A Tissues from mice This study N/A Chemicals, peptides, and recombinant proteins Luria-Bertani broth Qingdao Hope Cat# HB0128 Salmonella-Shigella Agar Qingdao Hope Cat# HB4089 Harmaline Macklin Cat# H832353 Mucin from porcine stomach Yuanye Cat# S12066 Metronidazole Yuanye Cat# S17079 Neomycin sulfate Rhawn Cat# R094126 Vancomycin hydrochloride Aladdin Cat# V105495 Ampicillin YuanYe Cat# S17018 Ceftiofur Sodium YuanYe Cat# S30940 L-cysteine Rhawn Cat# R014132 Ferrous sulfate Rhawn Cat# R010224 Iron (III) chloride YuanYe Cat# W12305 Sodium Tetrathionate Dihydrate Solarbio Cat# YS154920 HPF Probe Beyotime Cat# Y249872 DCFH-DA Beyotime Cat# S0033S Dihydroethidium (DHE) Beyotime Cat# S0064S Resazurin Beyotime Cat# Y025686 H2O2 Sigma-Aldrich Cat# 323381 TRIzol reagent Thermo Fisher Scientific Cat# 15596018CN SYTOTM MeilunBio Cat# MC7003 (Continued on next page) Cell Reports 45, 117858, August 25, 2026 19 ..

Control:

Mutagenesis:

Clinical Proteomics:

Recombinant:

Image Search Results

(A) HFF cells stably transduced with control sgRNA or sgRNA targeting UBE2I were infected with HSV-1 ICP0 RFm (MOI = 5). WCLs were analyzed by immunoblotting at 24 h post-infection, followed by densitometric quantification of band intensities. (B) HFF cells were infected with HSV-1 ICP0 RFm (MOI = 5) for 24 h. Denaturing immunoprecipitation was performed using anti-SUMO1 or anti-SUMO2/3 antibodies, and both input and immunoprecipitated samples were analyzed by immunoblotting. (C) U2OS cells stably transduced with control sgRNA or sgRNA targeting SUMO1 were infected with HSV-1 ΔICP0 (MOI = 5) for 24 h. WCLs were analyzed by immunoblotting, followed by densitometric quantification of band intensities. (D) U2OS cells stably transduced with control sgRNA or sgRNA targeting SUMO2 and SUMO3 were infected with HSV-1 ΔICP0 (MOI = 5) for 24 h. WCLs were analyzed by immunoblotting, followed by densitometric quantification of band intensities. (E) HEK293T cells were co-transfected with the indicated plasmids for 24 h and subsequently infected with HSV-1 ICP0 RFm (MOI = 5) for 24 h. Denaturing immunoprecipitation was performed, and both input and immunoprecipitated samples were analyzed by immunoblotting. (F) SMCHD1 -knockdown U2OS cells reconstituted with SMCHD1 WT, or the indicated mutants (Δhinge, K1848R, K1852R, K1872R, or the 3KR mutant [K1848R/K1852R/K1872R]) were infected with HSV-1 ΔICP0 (MOI = 5) for 24 h. WCLs were analyzed by immunoblotting, followed by densitometric quantification of band intensities. Data are presented as mean ± SD from three independent experiments (n = 3). Statistical significance was determined by one-way ANOVA or unpaired two tailed Student’s t test. p value: *, p < 0.05; **, p < 0.01; ***, p < 0.005. N.S.: no significance.

Journal: PLOS Pathogens

Article Title: SUMOylation and ubiquitination reciprocally regulate SMCHD1 antiviral activity against herpes simplex virus 1

doi: 10.1371/journal.ppat.1014371

Figure Lengend Snippet: (A) HFF cells stably transduced with control sgRNA or sgRNA targeting UBE2I were infected with HSV-1 ICP0 RFm (MOI = 5). WCLs were analyzed by immunoblotting at 24 h post-infection, followed by densitometric quantification of band intensities. (B) HFF cells were infected with HSV-1 ICP0 RFm (MOI = 5) for 24 h. Denaturing immunoprecipitation was performed using anti-SUMO1 or anti-SUMO2/3 antibodies, and both input and immunoprecipitated samples were analyzed by immunoblotting. (C) U2OS cells stably transduced with control sgRNA or sgRNA targeting SUMO1 were infected with HSV-1 ΔICP0 (MOI = 5) for 24 h. WCLs were analyzed by immunoblotting, followed by densitometric quantification of band intensities. (D) U2OS cells stably transduced with control sgRNA or sgRNA targeting SUMO2 and SUMO3 were infected with HSV-1 ΔICP0 (MOI = 5) for 24 h. WCLs were analyzed by immunoblotting, followed by densitometric quantification of band intensities. (E) HEK293T cells were co-transfected with the indicated plasmids for 24 h and subsequently infected with HSV-1 ICP0 RFm (MOI = 5) for 24 h. Denaturing immunoprecipitation was performed, and both input and immunoprecipitated samples were analyzed by immunoblotting. (F) SMCHD1 -knockdown U2OS cells reconstituted with SMCHD1 WT, or the indicated mutants (Δhinge, K1848R, K1852R, K1872R, or the 3KR mutant [K1848R/K1852R/K1872R]) were infected with HSV-1 ΔICP0 (MOI = 5) for 24 h. WCLs were analyzed by immunoblotting, followed by densitometric quantification of band intensities. Data are presented as mean ± SD from three independent experiments (n = 3). Statistical significance was determined by one-way ANOVA or unpaired two tailed Student’s t test. p value: *, p < 0.05; **, p < 0.01; ***, p < 0.005. N.S.: no significance.

Article Snippet: The diluted samples were incubated with anti-FLAG beads or with protein A/G beads together with rabbit control IgG (2 μg), anti-SUMO1 antibody (2 μg; A19121, Abclonal), or anti-SUMO2/3 antibody (2 μg; A22734, Abclonal) overnight at 4°C.

Techniques: Stable Transfection, Transduction, Control, Infection, Western Blot, Immunoprecipitation, Transfection, Knockdown, Mutagenesis, Two Tailed Test

a , Schematic overview of the HSV-1 ICP0 gene with mutations to disrupt either RNF8 binding site (T67A), E3 ligase function (C116G/C156A), Cyclin D3 binding site (D199A) or CoREST interaction (D671A/E673A). b , Representative immunofluorescence images of H4 cells co-transfected with TDP-43venus (green) and either vector, wild-type (WT) ICP0 or indicated ICP0 mutant. Cells were fixed 24 h post-transfection and stained for PML (white), ICP0 (red) and DAPI (blue). Scale bar, 10 µm. c , Quantification of nuclear TDP-43venus foci area (µm 2 ) (top) and PML foci count (bottom) of co-transfected cells in (b) using Foci Analyzer (ImageJ). Lines represent mean±SEM, dots represent individual cells (n = 37-74) of three independent biological replicates. *p<0.05, ***p<0.001 Kruskal-Wallis test followed by Dunn’s multiple comparisons test. d , Scatter plot of total nuclear TDP-43venus foci size (y-axis) against the nuclear PML dot count (x-axis). Each dot represents one corresponding measurement from (c). Blue dots represent cells overexpressing ICP0 WT, black dots represent cells transfected with vector. For association testing, all events were taken together in Spearman’s rank correlation test (r = –0.54, ***p<0.001, n = 118). e , Representative immunofluorescence images of H4 cells transduced with lentiviruses encoding TDP-43venus (green). After 3 days, cells were infected with HSV-1 and 1 h post-infection treated with proteasome inhibitor MG132 or DMSO, respectively. Cells were fixed 17 h post-infection and stained for PML (white), ICP27 (red) and DAPI (blue). Scale bar, 10 µm. f , Quantification of PML foci (top) and total nuclear TDP-43venus foci area (µm 2 ) (bottom) in (e). Lines represent mean±SEM. Dots represent individual cells, n = 58-362, three independent biological replicates, ***p<0.001 Kruskal-Wallis test followed by Dunn’s multiple comparisons test. g , Representative immunoblots showing SUMOylation status of FLAG-tagged TDP-43 purified by anti-FLAG immunoprecipitation (IP) from whole cell lysates (WCL) of HEK293T cells at 40 h post-transfection. Cells were co-transfected with HSV-1 ICP0 and/or stimulated with arsenite (500 µM, 1 h), as indicated. IP and WCL was analyzed by immunoblotting. Blots were stained with anti-SUMO2, anti-FLAG, anti-HSV-1 ICP0 and anti-GAPDH. h , Quantification of the full SUMO-2 lane in the TDP-43 IP of ( g ) normalized to the vector control. n = 3, **p<0.01 paired t test.

Journal: bioRxiv

Article Title: Herpes simplex virus infection promotes ALS pathology through ICP0-mediated PML body disruption

doi: 10.64898/2026.03.27.714707

Figure Lengend Snippet: a , Schematic overview of the HSV-1 ICP0 gene with mutations to disrupt either RNF8 binding site (T67A), E3 ligase function (C116G/C156A), Cyclin D3 binding site (D199A) or CoREST interaction (D671A/E673A). b , Representative immunofluorescence images of H4 cells co-transfected with TDP-43venus (green) and either vector, wild-type (WT) ICP0 or indicated ICP0 mutant. Cells were fixed 24 h post-transfection and stained for PML (white), ICP0 (red) and DAPI (blue). Scale bar, 10 µm. c , Quantification of nuclear TDP-43venus foci area (µm 2 ) (top) and PML foci count (bottom) of co-transfected cells in (b) using Foci Analyzer (ImageJ). Lines represent mean±SEM, dots represent individual cells (n = 37-74) of three independent biological replicates. *p<0.05, ***p<0.001 Kruskal-Wallis test followed by Dunn’s multiple comparisons test. d , Scatter plot of total nuclear TDP-43venus foci size (y-axis) against the nuclear PML dot count (x-axis). Each dot represents one corresponding measurement from (c). Blue dots represent cells overexpressing ICP0 WT, black dots represent cells transfected with vector. For association testing, all events were taken together in Spearman’s rank correlation test (r = –0.54, ***p<0.001, n = 118). e , Representative immunofluorescence images of H4 cells transduced with lentiviruses encoding TDP-43venus (green). After 3 days, cells were infected with HSV-1 and 1 h post-infection treated with proteasome inhibitor MG132 or DMSO, respectively. Cells were fixed 17 h post-infection and stained for PML (white), ICP27 (red) and DAPI (blue). Scale bar, 10 µm. f , Quantification of PML foci (top) and total nuclear TDP-43venus foci area (µm 2 ) (bottom) in (e). Lines represent mean±SEM. Dots represent individual cells, n = 58-362, three independent biological replicates, ***p<0.001 Kruskal-Wallis test followed by Dunn’s multiple comparisons test. g , Representative immunoblots showing SUMOylation status of FLAG-tagged TDP-43 purified by anti-FLAG immunoprecipitation (IP) from whole cell lysates (WCL) of HEK293T cells at 40 h post-transfection. Cells were co-transfected with HSV-1 ICP0 and/or stimulated with arsenite (500 µM, 1 h), as indicated. IP and WCL was analyzed by immunoblotting. Blots were stained with anti-SUMO2, anti-FLAG, anti-HSV-1 ICP0 and anti-GAPDH. h , Quantification of the full SUMO-2 lane in the TDP-43 IP of ( g ) normalized to the vector control. n = 3, **p<0.01 paired t test.

Article Snippet: Monoclonal mouse anti-FLAG-tag antibody, M2 (1:5,000), Sigma-Aldrich, F1804), anti-GAPDH (BioLegend, 607902; 1:1,000), Polyclonal rabbit anti-TDP-43 antibody (proteintech, 10782-2-AP, 1:2,000), mouse anti-lamin B (proteintech, 66095-1, 1:1,000), mouse anti-HSV-1 ICP0 (abcam, AB6513, 1:7,500), anti-SUMO2 (novus biologicals, NBP1-77163, 1:1,000) Secondary Antibodies: anti-rabbit IgG HRP (Promega, W401B, 1:10,000), anti-mouse IgG HRP (Promega, W402B, 1:10,000), Veriblot HRP (abcam, ab131366, 1:1,000)

Techniques: Binding Assay, Immunofluorescence, Transfection, Plasmid Preparation, Mutagenesis, Staining, Transduction, Infection, Western Blot, Purification, Immunoprecipitation, Control

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 Immunofluorescence images and quantification (n = 20) of SUMO1 (green) and SUMO2/3 (red) conjugation in RAW264.7 cells infected with STM 14028S over a 0-6 hrs time course. Scale bar, 10 µm. b Immunofluorescence images and quantification (n = 20) of SUMO1 (green) and SUMO2/3 (red) conjugation in RAW264.7 cells infected with STM WT or Δ ssaV or left uninfected (4 hpi). Scale bar, 10 µm. c Schematic of the high-content screening (HCS) workflow used to identify T3SS-2 effector(s) required for subversion of host SUMOylation. d SUMO2/3 suppression rates in RAW264.7 cells infected with STM WT or T3SS-2 effector knockout strains (4 hpi). For each sample, fluorescence intensity (FI) was measured across three fields (20 cells per field) to calculate the mean FI (MFI). Suppression rate = (MFI_uninfected - FI_test) / (MFI_uninfected - MFI_WT). e Representative immunofluorescence images of SUMO2/3 conjugation in RAW264.7 cells infected with STM WT, Δ ssaV , Δ sseK1 , or left uninfected (4 hpi). f Immunoblot analysis of SUMO2/3 conjugates in RAW264.7 cells infected with STM WT or Δ sseK1 or left uninfected (4 hpi). *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 infected with STM 14028S over a 0-6 hrs time course. Scale bar, 10 µm. b Immunofluorescence images and quantification (n = 20) of SUMO1 (green) and SUMO2/3 (red) conjugation in RAW264.7 cells infected with STM WT or Δ ssaV or left uninfected (4 hpi). Scale bar, 10 µm. c Schematic of the high-content screening (HCS) workflow used to identify T3SS-2 effector(s) required for subversion of host SUMOylation. d SUMO2/3 suppression rates in RAW264.7 cells infected with STM WT or T3SS-2 effector knockout strains (4 hpi). For each sample, fluorescence intensity (FI) was measured across three fields (20 cells per field) to calculate the mean FI (MFI). Suppression rate = (MFI_uninfected - FI_test) / (MFI_uninfected - MFI_WT). e Representative immunofluorescence images of SUMO2/3 conjugation in RAW264.7 cells infected with STM WT, Δ ssaV , Δ sseK1 , or left uninfected (4 hpi). f Immunoblot analysis of SUMO2/3 conjugates in RAW264.7 cells infected with STM WT or Δ sseK1 or left uninfected (4 hpi). *P < 0.05; ***P < 0.001; ns, not significant.

Techniques: Immunofluorescence, Conjugation Assay, Infection, High Content Screening, Knock-Out, Fluorescence, Western Blot

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

a Protein SUMOylation sites showing a >3-fold increase in SUMOylation intensity in STM Δ sseK1 -infected cells relative to WT. SUMOylation intensity was normalized to the corresponding protein abundance measured in proteomic datasets from STM Δ sseK1 - or WT-infected RAW264.7 cells. b GO and KEGG pathway enrichment analysis of proteins with SUMOylation intensity ratio (Δ sseK1 /WT) > 3. c, d IP analysis of Myd88 and Hspa8 SUMO2/3 modification in RAW264.7 cells infected with STM WT or Δ sseK1 or left uninfected (4 hpi). e Immunoblot analysis of SUMO2/3 modification in RAW-SseK1-OE and RAW-GFP-Ctrl cells. f Nine-quadrant plot showing protein abundance changes in two proteomic comparisons (RAW-SseK1-OE vs RAW-GFP-Ctrl and STM WT-infected vs Δ sseK1 -infected cells). Highlighted proteins are downregulated in both datasets. Fold-change values within dashed lines indicate <|1.3|. g Immunoblot analysis of PDCD4 expression in RAW-SseK1-OE and RAW-GFP-Ctrl cells. h Immunoblot analysis of PDCD4 and SUMO2/3 levels in RAW264.7 cells treated with increasing concentrations of the SUMOylation inhibitor 2-D08. ***P < 0.001.

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 Protein SUMOylation sites showing a >3-fold increase in SUMOylation intensity in STM Δ sseK1 -infected cells relative to WT. SUMOylation intensity was normalized to the corresponding protein abundance measured in proteomic datasets from STM Δ sseK1 - or WT-infected RAW264.7 cells. b GO and KEGG pathway enrichment analysis of proteins with SUMOylation intensity ratio (Δ sseK1 /WT) > 3. c, d IP analysis of Myd88 and Hspa8 SUMO2/3 modification in RAW264.7 cells infected with STM WT or Δ sseK1 or left uninfected (4 hpi). e Immunoblot analysis of SUMO2/3 modification in RAW-SseK1-OE and RAW-GFP-Ctrl cells. f Nine-quadrant plot showing protein abundance changes in two proteomic comparisons (RAW-SseK1-OE vs RAW-GFP-Ctrl and STM WT-infected vs Δ sseK1 -infected cells). Highlighted proteins are downregulated in both datasets. Fold-change values within dashed lines indicate <|1.3|. g Immunoblot analysis of PDCD4 expression in RAW-SseK1-OE and RAW-GFP-Ctrl cells. h Immunoblot analysis of PDCD4 and SUMO2/3 levels in RAW264.7 cells treated with increasing concentrations of the SUMOylation inhibitor 2-D08. ***P < 0.001.

Techniques: Infection, Quantitative Proteomics, Modification, Western Blot, Expressing

a Intracellular survival of STM WT and Δ sseK1 strains in RAW264.7 cells (1-6 hpi). n = 3. b Schematic of the experimental workflow for 2-D08 treatment and Salmonella infection of RAW264.7 cells. c Intracellular survival of STM WT and Δ sseK1 strains in RAW264.7 cells with or without 2-D08 pretreatment (4 hpi). d Schematic of the experimental workflow for 2-D08 administration and STM challenge in C57BL/6 mice. e Immunoblot analysis of SUMO2/3 modification in liver, spleen, and intestine tissues from C57BL/6 mice challenged with STM WT or Δ sseK1 , with or without 2-D08 pretreatment. f Bacterial load (CFU per gram of liver or spleen) in C57BL/6 mice infected with STM WT or Δ sseK1 . n = 4. g Survival curves of C57BL/6 mice challenged with STM WT or Δ sseK1 , with or without 2-D08 pretreatment. n = 6. *P < 0.05; **P < 0.01; ***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 Intracellular survival of STM WT and Δ sseK1 strains in RAW264.7 cells (1-6 hpi). n = 3. b Schematic of the experimental workflow for 2-D08 treatment and Salmonella infection of RAW264.7 cells. c Intracellular survival of STM WT and Δ sseK1 strains in RAW264.7 cells with or without 2-D08 pretreatment (4 hpi). d Schematic of the experimental workflow for 2-D08 administration and STM challenge in C57BL/6 mice. e Immunoblot analysis of SUMO2/3 modification in liver, spleen, and intestine tissues from C57BL/6 mice challenged with STM WT or Δ sseK1 , with or without 2-D08 pretreatment. f Bacterial load (CFU per gram of liver or spleen) in C57BL/6 mice infected with STM WT or Δ sseK1 . n = 4. g Survival curves of C57BL/6 mice challenged with STM WT or Δ sseK1 , with or without 2-D08 pretreatment. n = 6. *P < 0.05; **P < 0.01; ***P < 0.001; ns, not significant.

Techniques: Infection, Western Blot, Modification

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resource source identifier antibodies rabbit monoclonal anti sumo2 (ABclonal Biotechnology)

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