Review



otud1  (Bioss)


Bioz Verified Symbol Bioss is a verified supplier
Bioz Manufacturer Symbol Bioss manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
    • 94
      Buy from Supplier

    Structured Review

    Bioss otud1
    Endothelial <t>OTUD1</t> is significantly upregulated in diabetic wound tissues. (A) Real-time qPCR analysis for mRNA levels of OTU subfamily members in the skin wound tissues from control and T2DM mice (n = 7). (B) Immunoblotting and densitometric quantification analyses illustrating OTUD1 protein expression in skin wound tissues from both control and T2DM mice (n = 3). (C) Representative immunohistochemical images and (D) quantitative analysis of showing OTUD1-positive cells (brown) in the skin wound tissues of mice. Black arrowheads indicate positive OTUD1 signals (Scale bar = 50 μm; n = 7). (E) Immunoblot and quantitative analysis of OTUD1 protein expression in HUVECs, HaCaT, HDFa, and MPM cells (n = 4). (F) Representative immunofluorescence images displaying the colocalization of CD31 (red) and OTUD1 (green) in mouse skin wound tissues, with white arrowheads showing OTUD1 and CD31 colocalization sites. Tissues were counterstained with DAPI (blue; Scale bar = 50 μm). (G) Immunofluorescence staining and (H) quantitative analysis of OTUD1-positive cells (red) in both control and HG + PA-treated HUVECs for 4 h, counterstained with DAPI (blue; Scale bar = 50 μm). (I) Time-course study of OTUD1 expression in response to HG + PA in HUVECs, including immunoblot analysis and quantitative measurement (n = 3). Data are shown as mean ± SEM. Statistical analyses were performed using a two-tailed unpaired Student's t -test (A, B, D), Welch’s t test (H), and one-way ANOVA analysis followed by Bonferroni post-hoc test (E, I). HG + PA indicates treatment with 50 mM HG and 300 μM PA, unless specified otherwise. Abbreviations: Ctrl, control; T2DM, type 2 diabetes mellitus; OTU, ovarian tumor protease; HUVECs, human umbilical vein endothelial cells; HaCaT, human keratinocytes; HDFa, human dermal fibroblasts-adult; MPMs, mouse primary peritoneal macrophages; DAPI, 4ʹ,6-diamidino-2-phenylindole; HG + PA, high glucose plus palmitic acid; and OTUD1, ovarian tumor deubiquitinase 1.
    Otud1, supplied by Bioss, 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/result/otud1/product/Bioss
    Average 94 stars, based on 7 article reviews
    otud1 - by Bioz Stars, 2026-02
    94/100 stars

    Images

    1) Product Images from "OTUD1 delays wound healing by regulating endothelial function and angiogenesis in diabetic mice"

    Article Title: OTUD1 delays wound healing by regulating endothelial function and angiogenesis in diabetic mice

    Journal: Journal of Advanced Research

    doi: 10.1016/j.jare.2025.04.038

    Endothelial OTUD1 is significantly upregulated in diabetic wound tissues. (A) Real-time qPCR analysis for mRNA levels of OTU subfamily members in the skin wound tissues from control and T2DM mice (n = 7). (B) Immunoblotting and densitometric quantification analyses illustrating OTUD1 protein expression in skin wound tissues from both control and T2DM mice (n = 3). (C) Representative immunohistochemical images and (D) quantitative analysis of showing OTUD1-positive cells (brown) in the skin wound tissues of mice. Black arrowheads indicate positive OTUD1 signals (Scale bar = 50 μm; n = 7). (E) Immunoblot and quantitative analysis of OTUD1 protein expression in HUVECs, HaCaT, HDFa, and MPM cells (n = 4). (F) Representative immunofluorescence images displaying the colocalization of CD31 (red) and OTUD1 (green) in mouse skin wound tissues, with white arrowheads showing OTUD1 and CD31 colocalization sites. Tissues were counterstained with DAPI (blue; Scale bar = 50 μm). (G) Immunofluorescence staining and (H) quantitative analysis of OTUD1-positive cells (red) in both control and HG + PA-treated HUVECs for 4 h, counterstained with DAPI (blue; Scale bar = 50 μm). (I) Time-course study of OTUD1 expression in response to HG + PA in HUVECs, including immunoblot analysis and quantitative measurement (n = 3). Data are shown as mean ± SEM. Statistical analyses were performed using a two-tailed unpaired Student's t -test (A, B, D), Welch’s t test (H), and one-way ANOVA analysis followed by Bonferroni post-hoc test (E, I). HG + PA indicates treatment with 50 mM HG and 300 μM PA, unless specified otherwise. Abbreviations: Ctrl, control; T2DM, type 2 diabetes mellitus; OTU, ovarian tumor protease; HUVECs, human umbilical vein endothelial cells; HaCaT, human keratinocytes; HDFa, human dermal fibroblasts-adult; MPMs, mouse primary peritoneal macrophages; DAPI, 4ʹ,6-diamidino-2-phenylindole; HG + PA, high glucose plus palmitic acid; and OTUD1, ovarian tumor deubiquitinase 1.
    Figure Legend Snippet: Endothelial OTUD1 is significantly upregulated in diabetic wound tissues. (A) Real-time qPCR analysis for mRNA levels of OTU subfamily members in the skin wound tissues from control and T2DM mice (n = 7). (B) Immunoblotting and densitometric quantification analyses illustrating OTUD1 protein expression in skin wound tissues from both control and T2DM mice (n = 3). (C) Representative immunohistochemical images and (D) quantitative analysis of showing OTUD1-positive cells (brown) in the skin wound tissues of mice. Black arrowheads indicate positive OTUD1 signals (Scale bar = 50 μm; n = 7). (E) Immunoblot and quantitative analysis of OTUD1 protein expression in HUVECs, HaCaT, HDFa, and MPM cells (n = 4). (F) Representative immunofluorescence images displaying the colocalization of CD31 (red) and OTUD1 (green) in mouse skin wound tissues, with white arrowheads showing OTUD1 and CD31 colocalization sites. Tissues were counterstained with DAPI (blue; Scale bar = 50 μm). (G) Immunofluorescence staining and (H) quantitative analysis of OTUD1-positive cells (red) in both control and HG + PA-treated HUVECs for 4 h, counterstained with DAPI (blue; Scale bar = 50 μm). (I) Time-course study of OTUD1 expression in response to HG + PA in HUVECs, including immunoblot analysis and quantitative measurement (n = 3). Data are shown as mean ± SEM. Statistical analyses were performed using a two-tailed unpaired Student's t -test (A, B, D), Welch’s t test (H), and one-way ANOVA analysis followed by Bonferroni post-hoc test (E, I). HG + PA indicates treatment with 50 mM HG and 300 μM PA, unless specified otherwise. Abbreviations: Ctrl, control; T2DM, type 2 diabetes mellitus; OTU, ovarian tumor protease; HUVECs, human umbilical vein endothelial cells; HaCaT, human keratinocytes; HDFa, human dermal fibroblasts-adult; MPMs, mouse primary peritoneal macrophages; DAPI, 4ʹ,6-diamidino-2-phenylindole; HG + PA, high glucose plus palmitic acid; and OTUD1, ovarian tumor deubiquitinase 1.

    Techniques Used: Control, Western Blot, Expressing, Immunohistochemical staining, Immunofluorescence, Staining, Two Tailed Test

    OTUD1 deficiency rescues impaired wound healing by enhancing angiogenesis and fibrosis in T2DM mice. (A) Schematic diagram illustrating the animal experiment procedure. The mice (B) FBG levels and (C) body weight were recorded from weeks 9 to 16 (n = 7). * P < 0.05 vs WT-Sham; ** P < 0.01 vs WT-Sham; *** P < 0.001 vs WT-Sham; ns, no significance. ns (green) indicates that there is no statistical significance between OTUD1 −/− -T2DM and WT-T2DM. (D) Representative wound images and (E) wound closure rates are shown (n = 5). *** P < 0.001 vs WT-Sham; ## P < 0.01 vs WT-T2DM; ns, no significance. (F) H&E staining demonstrated regenerated skin at day 12 across different groups. Scale bar = 50 μm. (G) Quantitative assessments of epidermis thickness in mice (n = 7). (H) Masson's trichrome staining and (I) quantitative analysis of collagen deposition in skin wound tissues at day 12 (Scale bar = 50 μm; n = 7). (J) Representative images and (K) quantification of CD31-positive (brown) neovascularization via immunohistochemical staining at days 3, 7, and 12 (Scale bar = 50 μm; n = 7). Black arrows indicate skin neovascularization. (L-O) Immunoblotting and quantification of OTUD1, VEGFR2, p-eNOS, and eNOS in wound tissue lysates from Sham or T2DM mice with WT or OTUD1 knockout, normalized to GAPDH (n = 4). Data are displayed as mean ± SEM. Statistical analyses were performed using two-way ANOVA analysis followed by Bonferroni post-hoc test (B, C, E) and one-way ANOVA analysis followed by Bonferroni post-hoc test (G, I, K, M-O). Abbreviations: WT, wild-type; OTUD −/− , OTUD1-knockout; HFD, high-fat diet; STZ, streptozotocin; FBG, fasting blood glucose; eNOS, endothelial nitric oxide synthase.
    Figure Legend Snippet: OTUD1 deficiency rescues impaired wound healing by enhancing angiogenesis and fibrosis in T2DM mice. (A) Schematic diagram illustrating the animal experiment procedure. The mice (B) FBG levels and (C) body weight were recorded from weeks 9 to 16 (n = 7). * P < 0.05 vs WT-Sham; ** P < 0.01 vs WT-Sham; *** P < 0.001 vs WT-Sham; ns, no significance. ns (green) indicates that there is no statistical significance between OTUD1 −/− -T2DM and WT-T2DM. (D) Representative wound images and (E) wound closure rates are shown (n = 5). *** P < 0.001 vs WT-Sham; ## P < 0.01 vs WT-T2DM; ns, no significance. (F) H&E staining demonstrated regenerated skin at day 12 across different groups. Scale bar = 50 μm. (G) Quantitative assessments of epidermis thickness in mice (n = 7). (H) Masson's trichrome staining and (I) quantitative analysis of collagen deposition in skin wound tissues at day 12 (Scale bar = 50 μm; n = 7). (J) Representative images and (K) quantification of CD31-positive (brown) neovascularization via immunohistochemical staining at days 3, 7, and 12 (Scale bar = 50 μm; n = 7). Black arrows indicate skin neovascularization. (L-O) Immunoblotting and quantification of OTUD1, VEGFR2, p-eNOS, and eNOS in wound tissue lysates from Sham or T2DM mice with WT or OTUD1 knockout, normalized to GAPDH (n = 4). Data are displayed as mean ± SEM. Statistical analyses were performed using two-way ANOVA analysis followed by Bonferroni post-hoc test (B, C, E) and one-way ANOVA analysis followed by Bonferroni post-hoc test (G, I, K, M-O). Abbreviations: WT, wild-type; OTUD −/− , OTUD1-knockout; HFD, high-fat diet; STZ, streptozotocin; FBG, fasting blood glucose; eNOS, endothelial nitric oxide synthase.

    Techniques Used: Staining, Immunohistochemical staining, Western Blot, Knock-Out

    OTUD1 knockdown mitigates endothelial dysfunction and oxidative damage induced by HG + PA in vitro . HUVECs transfected with siOTUD1 or siNC were challenged with HG + PA. (A) Representative images and (B) quantitative analysis of scratch migration area in HUVECs at 0, 18, and 36 h with or without HG + PA treatment (n = 7). (C-F) Representative immunoblot and quantification of OTUD1, VEGFR2, p-eNOS, and eNOS with or without HG + PA exposure for 4 h (n = 4). Relative mRNA expression of (G) Vegfα , (H) Sdf1 , (I) Angpt1 , and (J) Tgfβ1 in HUVECs with or without HG + PA treatment for 24 h (n = 7). Flow cytometric analysis of (K, L) NO production and (M, N) ROS release in HUVECs with or without HG + PA treatment for 2 h (n = 7). Data are shown as mean ± SEM. Statistical analysis was conducted using one-way ANOVA analysis followed by Bonferroni post-hoc test (B, D-J, L, N). Abbreviations: NC, negative control; DAF-FM DA, 4-amino-5-methylamino-2′,7′-difluorofluorescein diacetate; DCFH-DA, 2′,7′-dichlorodihydrofluorescein diacetate; ROS, reactive oxygen species; NO, nitric oxide.
    Figure Legend Snippet: OTUD1 knockdown mitigates endothelial dysfunction and oxidative damage induced by HG + PA in vitro . HUVECs transfected with siOTUD1 or siNC were challenged with HG + PA. (A) Representative images and (B) quantitative analysis of scratch migration area in HUVECs at 0, 18, and 36 h with or without HG + PA treatment (n = 7). (C-F) Representative immunoblot and quantification of OTUD1, VEGFR2, p-eNOS, and eNOS with or without HG + PA exposure for 4 h (n = 4). Relative mRNA expression of (G) Vegfα , (H) Sdf1 , (I) Angpt1 , and (J) Tgfβ1 in HUVECs with or without HG + PA treatment for 24 h (n = 7). Flow cytometric analysis of (K, L) NO production and (M, N) ROS release in HUVECs with or without HG + PA treatment for 2 h (n = 7). Data are shown as mean ± SEM. Statistical analysis was conducted using one-way ANOVA analysis followed by Bonferroni post-hoc test (B, D-J, L, N). Abbreviations: NC, negative control; DAF-FM DA, 4-amino-5-methylamino-2′,7′-difluorofluorescein diacetate; DCFH-DA, 2′,7′-dichlorodihydrofluorescein diacetate; ROS, reactive oxygen species; NO, nitric oxide.

    Techniques Used: Knockdown, In Vitro, Transfection, Migration, Western Blot, Expressing, Negative Control

    OTUD1 directly interacts with β-catenin. (A) Schematic illustration of the quantitative proteomic screening used to identify OTUD1-binding proteins. Eleven potential binding partners were found, including CD44, BASP1, VARS1, LMAN1, GNG12, ITGA3, IARS1, DHX15, TMED10, JUP, and β-catenin. (B) HUVEC lysates were immunoprecipitated using OTUD1 or IgG antibodies, followed by immunoblotting with OTUD1 and β-catenin antibodies (n = 3). (C) Hek 293T cells were transfected for 24 h with Flag-vector or Flag-OTUD1 plasmids. Cell lysates underwent immunoprecipitation with anti-Flag or IgG antibodies, followed by immunoblotting using anti-Flag and β-catenin antibodies (n = 3). (D) BLI assay assessing β-catenin binding to in vitro -biotinylated OTUD1. Sensorgrams show the binding at different β-catenin concentrations (color lines). (E) Immunofluorescence staining for OTUD1 (red), β-catenin (green), and nuclei (DAPI, blue) in HUVECs. Scale bar = 50 μm. (F) Quantification of fluorescence intensity showing OTUD1 and β-catenin co-localization and (G) nuclear translocation of β-catenin (n = 7). White arrows indicate OTUD1 and β-catenin co-localization. (H) Diagram of the β-catenin domain deletion construct used in (I, J). (I, J) Full-length β-catenin or truncated forms were co-expressed with OTUD1 in Hek 293T cells. Immunoprecipitation was performed with anti-His antibody, followed by an immunoblot assay with specific antibodies (n = 3). Protein levels were normalized to GAPDH. Data are presented as mean ± SEM. Statistical analysis used a two-tailed unpaired Student's t -test (F, G). Abbreviations: IP, immunoprecipitation; IgG, immunoglobulin G; WCL, whole cell lysates; BLI, bio-layer interferometry; NTD, N-terminal domain; CTD, C-terminal domain; ARM, armadillo repeats 1–12.
    Figure Legend Snippet: OTUD1 directly interacts with β-catenin. (A) Schematic illustration of the quantitative proteomic screening used to identify OTUD1-binding proteins. Eleven potential binding partners were found, including CD44, BASP1, VARS1, LMAN1, GNG12, ITGA3, IARS1, DHX15, TMED10, JUP, and β-catenin. (B) HUVEC lysates were immunoprecipitated using OTUD1 or IgG antibodies, followed by immunoblotting with OTUD1 and β-catenin antibodies (n = 3). (C) Hek 293T cells were transfected for 24 h with Flag-vector or Flag-OTUD1 plasmids. Cell lysates underwent immunoprecipitation with anti-Flag or IgG antibodies, followed by immunoblotting using anti-Flag and β-catenin antibodies (n = 3). (D) BLI assay assessing β-catenin binding to in vitro -biotinylated OTUD1. Sensorgrams show the binding at different β-catenin concentrations (color lines). (E) Immunofluorescence staining for OTUD1 (red), β-catenin (green), and nuclei (DAPI, blue) in HUVECs. Scale bar = 50 μm. (F) Quantification of fluorescence intensity showing OTUD1 and β-catenin co-localization and (G) nuclear translocation of β-catenin (n = 7). White arrows indicate OTUD1 and β-catenin co-localization. (H) Diagram of the β-catenin domain deletion construct used in (I, J). (I, J) Full-length β-catenin or truncated forms were co-expressed with OTUD1 in Hek 293T cells. Immunoprecipitation was performed with anti-His antibody, followed by an immunoblot assay with specific antibodies (n = 3). Protein levels were normalized to GAPDH. Data are presented as mean ± SEM. Statistical analysis used a two-tailed unpaired Student's t -test (F, G). Abbreviations: IP, immunoprecipitation; IgG, immunoglobulin G; WCL, whole cell lysates; BLI, bio-layer interferometry; NTD, N-terminal domain; CTD, C-terminal domain; ARM, armadillo repeats 1–12.

    Techniques Used: Binding Assay, Immunoprecipitation, Western Blot, Transfection, Plasmid Preparation, In Vitro, Immunofluorescence, Staining, Fluorescence, Translocation Assay, Construct, Two Tailed Test

    OTUD1 modulates deubiquitinated modification of β-catenin. (A) Diagram showing the OTUD1 active site construct. (B) In vitro deubiquitylation assay of β-catenin by OTUD1. His-β-catenin and Myc-Ub were co-expressed with either Flag-OTUD1-WT or Flag-OTUD1-C320A in Hek 293T cells, followed by 10 μM MG132 treatment for 6 h. Ubiquitinated β-catenin was detected through immunoblotting (n = 3). (C) β-catenin ubiquitination assay in Hek 293T cells transfected with siOTUD1-1/2 and treated with MG132 for 6 h (n = 3). (D) Analysis of the ubiquitin chain type on β-catenin. His-β-catenin, Myc-Ub WT, Myc-Ub K48 only, Myc-Ub K48R, Myc-Ub K63 only, and Myc-Ub K63R were transfected into OTUD1 knockdown HUVECs, followed by MG132 treatment. Cell lysates were immunoprecipitated with anti-His antibodies and subsequent immunoblot analysis (n = 3). (E) OTUD1 overexpression diminished β-catenin K63 ubiquitination. His-β-catenin, Myc-Ub K63 only, and Flag-OTUD1 were co-expressed in Hek 293T cells, treated with MG132. Immunoprecipitation with anti-His antibodies was followed by immunoblotting (n = 3). (F) Identification of candidate ubiquitin sites on β-catenin. His-β-catenin WT or mutants, along with the Myc-Ub K63 plasmid, were transfected into HUVECs. Lysates underwent immunoprecipitation with anti-His antibodies and subjected to immunoblotting as indicated (n = 3). (G) OTUD1′s role in regulating β-catenin ubiquitination. His-β-catenin WT or 3KR, Myc-Ub K63 only, and Flag-OTUD1 were co-expressed in Hek 293T cells, followed by MG132 treatment. Cell lysates were immunoprecipitated with anti-His antibodies and analyzed by immunoblotting (n = 3). Abbreviations: C320A, cysteine at position 320 to alanine, MG132, Z-Leu-Leu-Leu-al; 3KR, lysine at position 496, 508, 625 to arginine.
    Figure Legend Snippet: OTUD1 modulates deubiquitinated modification of β-catenin. (A) Diagram showing the OTUD1 active site construct. (B) In vitro deubiquitylation assay of β-catenin by OTUD1. His-β-catenin and Myc-Ub were co-expressed with either Flag-OTUD1-WT or Flag-OTUD1-C320A in Hek 293T cells, followed by 10 μM MG132 treatment for 6 h. Ubiquitinated β-catenin was detected through immunoblotting (n = 3). (C) β-catenin ubiquitination assay in Hek 293T cells transfected with siOTUD1-1/2 and treated with MG132 for 6 h (n = 3). (D) Analysis of the ubiquitin chain type on β-catenin. His-β-catenin, Myc-Ub WT, Myc-Ub K48 only, Myc-Ub K48R, Myc-Ub K63 only, and Myc-Ub K63R were transfected into OTUD1 knockdown HUVECs, followed by MG132 treatment. Cell lysates were immunoprecipitated with anti-His antibodies and subsequent immunoblot analysis (n = 3). (E) OTUD1 overexpression diminished β-catenin K63 ubiquitination. His-β-catenin, Myc-Ub K63 only, and Flag-OTUD1 were co-expressed in Hek 293T cells, treated with MG132. Immunoprecipitation with anti-His antibodies was followed by immunoblotting (n = 3). (F) Identification of candidate ubiquitin sites on β-catenin. His-β-catenin WT or mutants, along with the Myc-Ub K63 plasmid, were transfected into HUVECs. Lysates underwent immunoprecipitation with anti-His antibodies and subjected to immunoblotting as indicated (n = 3). (G) OTUD1′s role in regulating β-catenin ubiquitination. His-β-catenin WT or 3KR, Myc-Ub K63 only, and Flag-OTUD1 were co-expressed in Hek 293T cells, followed by MG132 treatment. Cell lysates were immunoprecipitated with anti-His antibodies and analyzed by immunoblotting (n = 3). Abbreviations: C320A, cysteine at position 320 to alanine, MG132, Z-Leu-Leu-Leu-al; 3KR, lysine at position 496, 508, 625 to arginine.

    Techniques Used: Modification, Construct, In Vitro, Western Blot, Ubiquitin Proteomics, Transfection, Knockdown, Immunoprecipitation, Over Expression, Plasmid Preparation

    OTUD1 modulates β-catenin phosphorylation and nuclear translocation through deubiquitylation. (A) Representative western blot and (B) densitometric analysis of p-β-catenin, total-β-catenin, nuc-β-catenin, and cyt-β-catenin protein levels in skin wound tissues from different groups (n = 4). (C) Representative immunoblot analysis of β-catenin phosphorylation and nuclear translocation in different HUVECs groups, with (D) quantification of these protein levels (n = 4). (E) Immunoblot and (F) quantitative analysis of β-catenin phosphorylation and nuclear translocation in Hek 293T cells co-transfected with His-β-catenin WT or 3KR and Flag-OTUD1 (n = 3). (G) Representative images and (H) quantification of scratch migration area following co-expression of Flag-OTUD1 and His-β-catenin WT or 3KR in HUVECs at 0, 18, and 36 h (n = 7). Data are shown as mean ± SEM. Statistical analysis was performed using one-way ANOVA analysis followed by Bonferroni post-hoc test (B, D, F, H). Abbreviations: Nuc, nucleus; Cyt, cytoplasm.
    Figure Legend Snippet: OTUD1 modulates β-catenin phosphorylation and nuclear translocation through deubiquitylation. (A) Representative western blot and (B) densitometric analysis of p-β-catenin, total-β-catenin, nuc-β-catenin, and cyt-β-catenin protein levels in skin wound tissues from different groups (n = 4). (C) Representative immunoblot analysis of β-catenin phosphorylation and nuclear translocation in different HUVECs groups, with (D) quantification of these protein levels (n = 4). (E) Immunoblot and (F) quantitative analysis of β-catenin phosphorylation and nuclear translocation in Hek 293T cells co-transfected with His-β-catenin WT or 3KR and Flag-OTUD1 (n = 3). (G) Representative images and (H) quantification of scratch migration area following co-expression of Flag-OTUD1 and His-β-catenin WT or 3KR in HUVECs at 0, 18, and 36 h (n = 7). Data are shown as mean ± SEM. Statistical analysis was performed using one-way ANOVA analysis followed by Bonferroni post-hoc test (B, D, F, H). Abbreviations: Nuc, nucleus; Cyt, cytoplasm.

    Techniques Used: Phospho-proteomics, Translocation Assay, Western Blot, Transfection, Migration, Expressing

    Pharmacological inhibition of β-catenin reverses OTUD1 deletion-mediated recovery of delayed wound healing in db/db mice. (A) Schematic of the protocol for establishing the OTUD1 deletion mouse model in db/m or db/db mice. The mice (B) FBG levels and (C) body weight of the indicated mice at weeks 10, 12, 14, 16, 18, 20, 22, and 24 (n = 7). *** P < 0.001 vs db/m -AAV-shNC. ns (green) indicates that there is no statistical significance between db/db -AAV-shNC and db/db -AAV-shOTUD1. ns (purple) indicates that there is no statistical significance between db/db -AAV-shOTUD1 and db/db -AAV-shOTUD1-MSAB. (D) Skin wound healing images and (E) statistical analysis of wound closure rate (n = 5). ** P < 0.01 vs db/m -AAV-shNC; *** P < 0.001 vs db/m -AAV-shNC; ## P < 0.01 vs db/db -AAV-shNC; ### P < 0.001 vs db/db -AAV-shNC; && P < 0.01 vs db/db -AAV-shOTUD1; ns, no significance. (F) H&E staining images of skin wound tissues. Scale bar = 50 μm. (G) Representative Masson’s trichrome staining in skin wound tissues. Scale bar = 50 μm. (H) CD31 immunohistochemical staining of neovascularization in skin wound tissues on days 3, 7, and 12 (Scale bar = 50 μm). The black arrows denote skin neovascularization. (I) Representative immunoblotting and (J-M) quantitative analysis of OTUD1, VEGFR2, Nuc-β-catenin, p-eNOS, and eNOS protein levels in skin wound tissues from different groups (n = 4). Data are shown as mean ± SEM. Statistical analyses were performed using two-way ANOVA analysis followed by Bonferroni post-hoc test (B, C, E) and one-way ANOVA analysis followed by Bonferroni post-hoc test (J-M). Abbreviations: MSAB, ethionine sulfoxide β-methyl ester; AAV2/BI30-shOTUD1, adeno-associated virus serotype 2/BI30 carrying shOTUD1 under the CMV promoter.
    Figure Legend Snippet: Pharmacological inhibition of β-catenin reverses OTUD1 deletion-mediated recovery of delayed wound healing in db/db mice. (A) Schematic of the protocol for establishing the OTUD1 deletion mouse model in db/m or db/db mice. The mice (B) FBG levels and (C) body weight of the indicated mice at weeks 10, 12, 14, 16, 18, 20, 22, and 24 (n = 7). *** P < 0.001 vs db/m -AAV-shNC. ns (green) indicates that there is no statistical significance between db/db -AAV-shNC and db/db -AAV-shOTUD1. ns (purple) indicates that there is no statistical significance between db/db -AAV-shOTUD1 and db/db -AAV-shOTUD1-MSAB. (D) Skin wound healing images and (E) statistical analysis of wound closure rate (n = 5). ** P < 0.01 vs db/m -AAV-shNC; *** P < 0.001 vs db/m -AAV-shNC; ## P < 0.01 vs db/db -AAV-shNC; ### P < 0.001 vs db/db -AAV-shNC; && P < 0.01 vs db/db -AAV-shOTUD1; ns, no significance. (F) H&E staining images of skin wound tissues. Scale bar = 50 μm. (G) Representative Masson’s trichrome staining in skin wound tissues. Scale bar = 50 μm. (H) CD31 immunohistochemical staining of neovascularization in skin wound tissues on days 3, 7, and 12 (Scale bar = 50 μm). The black arrows denote skin neovascularization. (I) Representative immunoblotting and (J-M) quantitative analysis of OTUD1, VEGFR2, Nuc-β-catenin, p-eNOS, and eNOS protein levels in skin wound tissues from different groups (n = 4). Data are shown as mean ± SEM. Statistical analyses were performed using two-way ANOVA analysis followed by Bonferroni post-hoc test (B, C, E) and one-way ANOVA analysis followed by Bonferroni post-hoc test (J-M). Abbreviations: MSAB, ethionine sulfoxide β-methyl ester; AAV2/BI30-shOTUD1, adeno-associated virus serotype 2/BI30 carrying shOTUD1 under the CMV promoter.

    Techniques Used: Inhibition, Staining, Immunohistochemical staining, Western Blot, Virus

    OTUD1 knockdown mitigates HG + PA-induced endothelial dysfunction via β-catenin translocation. HUVECs transfected with siOTUD1 or siNC were exposed with HG + PA. Cells were pretreated with MSAB (10 μM) for 1 h, while DMSO was used at the same concentration as a control. The migration capacity of HUVECs was evaluated using an in vitro scratch wound assay. (A) Representative images at specified time points. (B) Percentage of wound migration area (n = 7). (C-G) Representative immunoblot and quantitative analysis of OTUD1, VEGFR2, p-eNOS, eNOS, and Nuc-β-catenin in HUVECs with HG + PA treatment for 4 h (n = 4). Relative mRNA expression levels of (H) Vegfα , (I) Sdf1 , (J) Angpt1 , and (K) Tgfβ1 in HUVECs across different groups following HG + PA treatment for 24 h (n = 7). (L and M) NO release measured via flow cytometry using the DAF-FM DA probe after HG + PA exposure for 2 h, followed by quantification (n = 7). (N and O) ROS production assessed by flow cytometry with the DCFH-DA kit with HG + PA exposure for 2 h and quantified (n = 7). Data are shown as mean ± SEM. Statistical analyses were performed using one-way ANOVA analysis followed by Bonferroni post-hoc test (B, D–K, M) and Kruskal–Wallis test with Dunn’s post hoc test (O). Abbreviations: DMSO, dimethyl sulfoxide.
    Figure Legend Snippet: OTUD1 knockdown mitigates HG + PA-induced endothelial dysfunction via β-catenin translocation. HUVECs transfected with siOTUD1 or siNC were exposed with HG + PA. Cells were pretreated with MSAB (10 μM) for 1 h, while DMSO was used at the same concentration as a control. The migration capacity of HUVECs was evaluated using an in vitro scratch wound assay. (A) Representative images at specified time points. (B) Percentage of wound migration area (n = 7). (C-G) Representative immunoblot and quantitative analysis of OTUD1, VEGFR2, p-eNOS, eNOS, and Nuc-β-catenin in HUVECs with HG + PA treatment for 4 h (n = 4). Relative mRNA expression levels of (H) Vegfα , (I) Sdf1 , (J) Angpt1 , and (K) Tgfβ1 in HUVECs across different groups following HG + PA treatment for 24 h (n = 7). (L and M) NO release measured via flow cytometry using the DAF-FM DA probe after HG + PA exposure for 2 h, followed by quantification (n = 7). (N and O) ROS production assessed by flow cytometry with the DCFH-DA kit with HG + PA exposure for 2 h and quantified (n = 7). Data are shown as mean ± SEM. Statistical analyses were performed using one-way ANOVA analysis followed by Bonferroni post-hoc test (B, D–K, M) and Kruskal–Wallis test with Dunn’s post hoc test (O). Abbreviations: DMSO, dimethyl sulfoxide.

    Techniques Used: Knockdown, Translocation Assay, Transfection, Concentration Assay, Control, Migration, In Vitro, Scratch Wound Assay Assay, Western Blot, Expressing, Flow Cytometry



    Similar Products

    Image Search Results