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monoclonal antibody against mouse fzd4  (R&D Systems)


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    R&D Systems monoclonal antibody against mouse fzd4
    <t>FZD4</t> knockout mice develop FEVR features. FZD4 knockout mice (FZD4KO) was used as an FEVR model. (A) Western blot for FZD4 in the retinal tissue of FZD4 knockout mice, compared to wildtype controls (WT). (B–C) Electroretinogram (ERG) assessments at 16 weeks, shown by representative chars (B) and by quantification (C). Blue circle: b-wave; green circle: a-wave. (D) Retinal vascular density by CD31 + area quantification. N = 5. ∗p < 0.05. NS: no significant.
    Monoclonal Antibody Against Mouse Fzd4, supplied by R&D Systems, used in various techniques. Bioz Stars score: 94/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/anti+fzd4/Human+CCR6+Antibody/pmc12750777-33-18-24
    Average 94 stars, based on 10 article reviews
    monoclonal antibody against mouse fzd4 - by Bioz Stars, 2026-09
    94/100 stars

    Images

    1) Product Images from "Comparative analysis of activation of macrophages/microglia in diabetic retinopathy and Familial Exudative Vitreoretinopathy"

    Article Title: Comparative analysis of activation of macrophages/microglia in diabetic retinopathy and Familial Exudative Vitreoretinopathy

    Journal: Biochemistry and Biophysics Reports

    doi: 10.1016/j.bbrep.2025.102396

    FZD4 knockout mice develop FEVR features. FZD4 knockout mice (FZD4KO) was used as an FEVR model. (A) Western blot for FZD4 in the retinal tissue of FZD4 knockout mice, compared to wildtype controls (WT). (B–C) Electroretinogram (ERG) assessments at 16 weeks, shown by representative chars (B) and by quantification (C). Blue circle: b-wave; green circle: a-wave. (D) Retinal vascular density by CD31 + area quantification. N = 5. ∗p < 0.05. NS: no significant.
    Figure Legend Snippet: FZD4 knockout mice develop FEVR features. FZD4 knockout mice (FZD4KO) was used as an FEVR model. (A) Western blot for FZD4 in the retinal tissue of FZD4 knockout mice, compared to wildtype controls (WT). (B–C) Electroretinogram (ERG) assessments at 16 weeks, shown by representative chars (B) and by quantification (C). Blue circle: b-wave; green circle: a-wave. (D) Retinal vascular density by CD31 + area quantification. N = 5. ∗p < 0.05. NS: no significant.

    Techniques Used: Knock-Out, Western Blot

    Related Articles

    other:

    Article Title: The Lin28b/Wnt5a axis drives pancreas cancer through crosstalk between cancer associated fibroblasts and tumor epithelium.
    Article Snippet: Cells were sorted on the FACSAriaTM II cell sorter (BD) for CD45/CD31/ EpCAM- PDPN+ Fzd4+ and CD45/CD31/EpCAM- PDPN+ Fzd4- cell populations.

    Western Blot:

    Article Title: The Lin28b/Wnt5a axis drives pancreas cancer through crosstalk between cancer associated fibroblasts and tumor epithelium
    Article Snippet: .. The antibodies used were anti-Lin28B (Abcam, #ab191881, 1:1000; abcepta, #AP1485C, 1:1000), anti-actin (Santa Cruz, #sc-7210, 1:1000), anti-Wnt5a (Abcam, #ab229200, 1:500; R&D, #MAB645, 1:200), anti-Wnt10a (ABclonal, #A15602, 1:1000; biodragon, #BD-PNO283, 1:1000), anti-β-catenin (Abcam, #ab32572, 1:10,000), anti-Pcsk9 (Abcam, #ab185194, 1:1000), anti-PGRN (Abcam, #ab187070, 1:1000), and anti- Ldlr (Abcam, #ab52818, 1:500) for WB; anti-Ki67 (Abcam, #ab16667, 0.03 μg/ml), anti-α-SMA (Gene Tech, #GM085129, 1:100), anti-CK19 (Proteintech, #10712-1-AP, 0.5 μg/ml), anti-Lin28B (Abcam, #ab191881, 1:50; abcepta, #AP1485C, 1:50) and anti-Pcsk9 (Abcam, #ab185194, 1:100) for IF and IHC; anti-Fzd4 (R&D Systems, #MAB194, 1:100), rabbit anti-rat IgG-PE (Solarbio, #K0032R-PE, 1:100), anti-CD45-AlexaFluor 647 (BioLegend, #103124, 1:200), anti-CD326 (Ep- CAM)- AlexaFluor 488 (BioLegend, #118212, 1:25), anti-CD31-AlexaFluor 647 (BioLegend, #102416, 1:200) and anti-PDPN-APC/Cy7 (BioLegend, #127418, 1:200) for FACS. .. Recombinant protein of Wnt5a (#APP549Mu01) was purchased from Cloud-Clone.

    Article Title: The Lin28b/Wnt5a axis drives pancreas cancer through crosstalk between cancer associated fibroblasts and tumor epithelium.
    Article Snippet: Recombinant protein of Wnt5a (#APP549Mu01) was purchased from Cloud-Clone. .. The antibodies used were anti-Lin28B (Abcam, #ab191881, 1:1000; abcepta, #AP1485C, 1:1000), anti-actin (Santa Cruz, #sc-7210, 1:1000), anti-Wnt5a (Abcam, #ab229200, 1:500; R&D, #MAB645, 1:200), antiWnt10a (ABclonal, #A15602, 1:1000; biodragon, #BD-PNO283, 1:1000), anti-β-catenin (Abcam, #ab32572, 1:10,000), anti-Pcsk9 (Abcam, #ab185194, 1:1000), anti-PGRN (Abcam, #ab187070, 1:1000), and antiLdlr (Abcam, #ab52818, 1:500) for WB; anti-Ki67 (Abcam, #ab16667, 0.03μg/ml), anti-α-SMA (Gene Tech, #GM085129, 1:100), anti-CK19 (Proteintech, #10712-1-AP, 0.5 μg/ml), anti-Lin28B (Abcam, #ab191881, 1:50; abcepta, #AP1485C, 1:50) and anti-Pcsk9 (Abcam, #ab185194, 1:100) for IF and IHC; anti-Fzd4 (R&D Systems, #MAB194, 1:100), rabbit anti-rat IgG-PE (Solarbio, #K0032R-PE, 1:100), anti-CD45-AlexaFluor 647 (BioLegend, #103124, 1:200), anti-CD326 (Ep-CAM)-AlexaFluor 488 (BioLegend, #118212, 1:25), anti-CD31-AlexaFluor 647 (BioLegend, #102416, 1:200) and anti-PDPN-APC/Cy7 (BioLegend, #127418, 1:200) for FACS. .. Doxycycline (#S5159) was purchased from Selleck.

    Article Title: Wnt- β -Catenin Signaling Promotes the Maturation of Mast Cells
    Article Snippet: The lysates were subjected to SDS-PAGE on 7.5% polyacrylamide gel and were then transferred onto polyvinylidene fluoride membranes (Millipore). .. After the reaction was blocked with 5% skim milk in TBS containing 0.1% Tween 20 at room temperature for 1 h, the membranes were incubated with mouse anti- β -catenin (Santa Cruz Biotechnology), anti-LRP5 (Santa Cruz Biotechnology), anti-Fzd4 (R&D Systems) or mouse anti- β -actin (Sigma-Aldrich) antibodies at 4°C overnight, followed by reaction with HRP-conjugated anti-mouse IgG antibody (Cell signaling Technology) at room temperature for 1 h. The bands were visualized by ECL Plus Western blotting detection regents (GE Healthcare) and the signals were read using a LAS-4000 imaging system (FUJI Film). .. BMMCs were cocultured with mitomycin C-treated Swiss 3T3 fibroblasts in the presence of 100 ng/mL SCF and 1 μ g/mL normal rat IgG (R&D Systems) or 1 μ g/mL anti-Wnt5a antibody (R&D Systems).

    Immunohistochemistry:

    Article Title: The Lin28b/Wnt5a axis drives pancreas cancer through crosstalk between cancer associated fibroblasts and tumor epithelium
    Article Snippet: .. The antibodies used were anti-Lin28B (Abcam, #ab191881, 1:1000; abcepta, #AP1485C, 1:1000), anti-actin (Santa Cruz, #sc-7210, 1:1000), anti-Wnt5a (Abcam, #ab229200, 1:500; R&D, #MAB645, 1:200), anti-Wnt10a (ABclonal, #A15602, 1:1000; biodragon, #BD-PNO283, 1:1000), anti-β-catenin (Abcam, #ab32572, 1:10,000), anti-Pcsk9 (Abcam, #ab185194, 1:1000), anti-PGRN (Abcam, #ab187070, 1:1000), and anti- Ldlr (Abcam, #ab52818, 1:500) for WB; anti-Ki67 (Abcam, #ab16667, 0.03 μg/ml), anti-α-SMA (Gene Tech, #GM085129, 1:100), anti-CK19 (Proteintech, #10712-1-AP, 0.5 μg/ml), anti-Lin28B (Abcam, #ab191881, 1:50; abcepta, #AP1485C, 1:50) and anti-Pcsk9 (Abcam, #ab185194, 1:100) for IF and IHC; anti-Fzd4 (R&D Systems, #MAB194, 1:100), rabbit anti-rat IgG-PE (Solarbio, #K0032R-PE, 1:100), anti-CD45-AlexaFluor 647 (BioLegend, #103124, 1:200), anti-CD326 (Ep- CAM)- AlexaFluor 488 (BioLegend, #118212, 1:25), anti-CD31-AlexaFluor 647 (BioLegend, #102416, 1:200) and anti-PDPN-APC/Cy7 (BioLegend, #127418, 1:200) for FACS. .. Recombinant protein of Wnt5a (#APP549Mu01) was purchased from Cloud-Clone.

    Article Title: The Lin28b/Wnt5a axis drives pancreas cancer through crosstalk between cancer associated fibroblasts and tumor epithelium.
    Article Snippet: Recombinant protein of Wnt5a (#APP549Mu01) was purchased from Cloud-Clone. .. The antibodies used were anti-Lin28B (Abcam, #ab191881, 1:1000; abcepta, #AP1485C, 1:1000), anti-actin (Santa Cruz, #sc-7210, 1:1000), anti-Wnt5a (Abcam, #ab229200, 1:500; R&D, #MAB645, 1:200), antiWnt10a (ABclonal, #A15602, 1:1000; biodragon, #BD-PNO283, 1:1000), anti-β-catenin (Abcam, #ab32572, 1:10,000), anti-Pcsk9 (Abcam, #ab185194, 1:1000), anti-PGRN (Abcam, #ab187070, 1:1000), and antiLdlr (Abcam, #ab52818, 1:500) for WB; anti-Ki67 (Abcam, #ab16667, 0.03μg/ml), anti-α-SMA (Gene Tech, #GM085129, 1:100), anti-CK19 (Proteintech, #10712-1-AP, 0.5 μg/ml), anti-Lin28B (Abcam, #ab191881, 1:50; abcepta, #AP1485C, 1:50) and anti-Pcsk9 (Abcam, #ab185194, 1:100) for IF and IHC; anti-Fzd4 (R&D Systems, #MAB194, 1:100), rabbit anti-rat IgG-PE (Solarbio, #K0032R-PE, 1:100), anti-CD45-AlexaFluor 647 (BioLegend, #103124, 1:200), anti-CD326 (Ep-CAM)-AlexaFluor 488 (BioLegend, #118212, 1:25), anti-CD31-AlexaFluor 647 (BioLegend, #102416, 1:200) and anti-PDPN-APC/Cy7 (BioLegend, #127418, 1:200) for FACS. .. Doxycycline (#S5159) was purchased from Selleck.

    Chick Chorioallantoic Membrane Assay:

    Article Title: The Lin28b/Wnt5a axis drives pancreas cancer through crosstalk between cancer associated fibroblasts and tumor epithelium
    Article Snippet: .. The antibodies used were anti-Lin28B (Abcam, #ab191881, 1:1000; abcepta, #AP1485C, 1:1000), anti-actin (Santa Cruz, #sc-7210, 1:1000), anti-Wnt5a (Abcam, #ab229200, 1:500; R&D, #MAB645, 1:200), anti-Wnt10a (ABclonal, #A15602, 1:1000; biodragon, #BD-PNO283, 1:1000), anti-β-catenin (Abcam, #ab32572, 1:10,000), anti-Pcsk9 (Abcam, #ab185194, 1:1000), anti-PGRN (Abcam, #ab187070, 1:1000), and anti- Ldlr (Abcam, #ab52818, 1:500) for WB; anti-Ki67 (Abcam, #ab16667, 0.03 μg/ml), anti-α-SMA (Gene Tech, #GM085129, 1:100), anti-CK19 (Proteintech, #10712-1-AP, 0.5 μg/ml), anti-Lin28B (Abcam, #ab191881, 1:50; abcepta, #AP1485C, 1:50) and anti-Pcsk9 (Abcam, #ab185194, 1:100) for IF and IHC; anti-Fzd4 (R&D Systems, #MAB194, 1:100), rabbit anti-rat IgG-PE (Solarbio, #K0032R-PE, 1:100), anti-CD45-AlexaFluor 647 (BioLegend, #103124, 1:200), anti-CD326 (Ep- CAM)- AlexaFluor 488 (BioLegend, #118212, 1:25), anti-CD31-AlexaFluor 647 (BioLegend, #102416, 1:200) and anti-PDPN-APC/Cy7 (BioLegend, #127418, 1:200) for FACS. .. Recombinant protein of Wnt5a (#APP549Mu01) was purchased from Cloud-Clone.

    FACS:

    Article Title: The Lin28b/Wnt5a axis drives pancreas cancer through crosstalk between cancer associated fibroblasts and tumor epithelium
    Article Snippet: .. The antibodies used were anti-Lin28B (Abcam, #ab191881, 1:1000; abcepta, #AP1485C, 1:1000), anti-actin (Santa Cruz, #sc-7210, 1:1000), anti-Wnt5a (Abcam, #ab229200, 1:500; R&D, #MAB645, 1:200), anti-Wnt10a (ABclonal, #A15602, 1:1000; biodragon, #BD-PNO283, 1:1000), anti-β-catenin (Abcam, #ab32572, 1:10,000), anti-Pcsk9 (Abcam, #ab185194, 1:1000), anti-PGRN (Abcam, #ab187070, 1:1000), and anti- Ldlr (Abcam, #ab52818, 1:500) for WB; anti-Ki67 (Abcam, #ab16667, 0.03 μg/ml), anti-α-SMA (Gene Tech, #GM085129, 1:100), anti-CK19 (Proteintech, #10712-1-AP, 0.5 μg/ml), anti-Lin28B (Abcam, #ab191881, 1:50; abcepta, #AP1485C, 1:50) and anti-Pcsk9 (Abcam, #ab185194, 1:100) for IF and IHC; anti-Fzd4 (R&D Systems, #MAB194, 1:100), rabbit anti-rat IgG-PE (Solarbio, #K0032R-PE, 1:100), anti-CD45-AlexaFluor 647 (BioLegend, #103124, 1:200), anti-CD326 (Ep- CAM)- AlexaFluor 488 (BioLegend, #118212, 1:25), anti-CD31-AlexaFluor 647 (BioLegend, #102416, 1:200) and anti-PDPN-APC/Cy7 (BioLegend, #127418, 1:200) for FACS. .. Recombinant protein of Wnt5a (#APP549Mu01) was purchased from Cloud-Clone.

    Article Title: The Lin28b/Wnt5a axis drives pancreas cancer through crosstalk between cancer associated fibroblasts and tumor epithelium.
    Article Snippet: Recombinant protein of Wnt5a (#APP549Mu01) was purchased from Cloud-Clone. .. The antibodies used were anti-Lin28B (Abcam, #ab191881, 1:1000; abcepta, #AP1485C, 1:1000), anti-actin (Santa Cruz, #sc-7210, 1:1000), anti-Wnt5a (Abcam, #ab229200, 1:500; R&D, #MAB645, 1:200), antiWnt10a (ABclonal, #A15602, 1:1000; biodragon, #BD-PNO283, 1:1000), anti-β-catenin (Abcam, #ab32572, 1:10,000), anti-Pcsk9 (Abcam, #ab185194, 1:1000), anti-PGRN (Abcam, #ab187070, 1:1000), and antiLdlr (Abcam, #ab52818, 1:500) for WB; anti-Ki67 (Abcam, #ab16667, 0.03μg/ml), anti-α-SMA (Gene Tech, #GM085129, 1:100), anti-CK19 (Proteintech, #10712-1-AP, 0.5 μg/ml), anti-Lin28B (Abcam, #ab191881, 1:50; abcepta, #AP1485C, 1:50) and anti-Pcsk9 (Abcam, #ab185194, 1:100) for IF and IHC; anti-Fzd4 (R&D Systems, #MAB194, 1:100), rabbit anti-rat IgG-PE (Solarbio, #K0032R-PE, 1:100), anti-CD45-AlexaFluor 647 (BioLegend, #103124, 1:200), anti-CD326 (Ep-CAM)-AlexaFluor 488 (BioLegend, #118212, 1:25), anti-CD31-AlexaFluor 647 (BioLegend, #102416, 1:200) and anti-PDPN-APC/Cy7 (BioLegend, #127418, 1:200) for FACS. .. Doxycycline (#S5159) was purchased from Selleck.

    Incubation:

    Article Title: Wnt- β -Catenin Signaling Promotes the Maturation of Mast Cells
    Article Snippet: The lysates were subjected to SDS-PAGE on 7.5% polyacrylamide gel and were then transferred onto polyvinylidene fluoride membranes (Millipore). .. After the reaction was blocked with 5% skim milk in TBS containing 0.1% Tween 20 at room temperature for 1 h, the membranes were incubated with mouse anti- β -catenin (Santa Cruz Biotechnology), anti-LRP5 (Santa Cruz Biotechnology), anti-Fzd4 (R&D Systems) or mouse anti- β -actin (Sigma-Aldrich) antibodies at 4°C overnight, followed by reaction with HRP-conjugated anti-mouse IgG antibody (Cell signaling Technology) at room temperature for 1 h. The bands were visualized by ECL Plus Western blotting detection regents (GE Healthcare) and the signals were read using a LAS-4000 imaging system (FUJI Film). .. BMMCs were cocultured with mitomycin C-treated Swiss 3T3 fibroblasts in the presence of 100 ng/mL SCF and 1 μ g/mL normal rat IgG (R&D Systems) or 1 μ g/mL anti-Wnt5a antibody (R&D Systems).

    Imaging:

    Article Title: Wnt- β -Catenin Signaling Promotes the Maturation of Mast Cells
    Article Snippet: The lysates were subjected to SDS-PAGE on 7.5% polyacrylamide gel and were then transferred onto polyvinylidene fluoride membranes (Millipore). .. After the reaction was blocked with 5% skim milk in TBS containing 0.1% Tween 20 at room temperature for 1 h, the membranes were incubated with mouse anti- β -catenin (Santa Cruz Biotechnology), anti-LRP5 (Santa Cruz Biotechnology), anti-Fzd4 (R&D Systems) or mouse anti- β -actin (Sigma-Aldrich) antibodies at 4°C overnight, followed by reaction with HRP-conjugated anti-mouse IgG antibody (Cell signaling Technology) at room temperature for 1 h. The bands were visualized by ECL Plus Western blotting detection regents (GE Healthcare) and the signals were read using a LAS-4000 imaging system (FUJI Film). .. BMMCs were cocultured with mitomycin C-treated Swiss 3T3 fibroblasts in the presence of 100 ng/mL SCF and 1 μ g/mL normal rat IgG (R&D Systems) or 1 μ g/mL anti-Wnt5a antibody (R&D Systems).

    Staining:

    Article Title: The Lin28b/Wnt5a axis drives pancreas cancer through crosstalk between cancer associated fibroblasts and tumor epithelium
    Article Snippet: .. Cells were stained with anti-Fzd4 (R&D Systems, #MAB194), and rabbit anti-rat IgG-PE (Solarbio, #K0032R-PE) was used as the secondary antibody. .. Cells were subsequently stained with anti-CD45-AlexaFluor 647 (BioLegend, #103124), anti-CD326 (Ep-CAM)-AlexaFluor 488 (BioLegend, #118212), anti-CD31-AlexaFluor 647 (BioLegend, #102416) and anti-PDPN-APC/Cy7 (BioLegend, #127418).



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    ABclonal Biotechnology rabbit anti-fzd4 a8161
    Rnf43 regulates the level of TJ through <t>Fzd4.</t> A) Immunofluorescent staining for Zo‐1 with bEnd.3 lacking <t>Fzd4,</t> Fzd6 or Fzd10. Scale bar, 50 µm. B) WB detection for TJ proteins and β‐catenin with bEnd.3 lacking Fzd4, Fzd6 or Fzd10. C) Statistic analysis of relative intensity of TJ proteins and β‐catenin in WB detection, n=4. D) Co‐IP detection of Rnf43 and Fzd4. E) Co‐location detection of Rnf43 and Fzd4. Scale bar, 10 µm. F) WB detection for Fzd4 after Rnf43 knockdown. G) Statistic analysis of relative intensity of Fzd4 in WB detection, n=4. H) Detection of Fzd4 after Rnf43 knockdown. Scale bar, 10 µm. I) Statistic analysis of relative signal intensity of Fzd4, n=4. Data were presented as mean ± SEM, one‐way ANOVA, Two‐tailed Student's t ‐test, ns, no significant difference, * p < 0.05, ** p < 0.01, *** p < 0.001.
    Rabbit Anti Fzd4 A8161, supplied by ABclonal Biotechnology, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/anti+fzd4/fzd4+antibody/pm37554197-47-17-20
    Average 90 stars, based on 1 article reviews
    rabbit anti-fzd4 a8161 - by Bioz Stars, 2026-09
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    FZD4 knockout mice develop FEVR features. FZD4 knockout mice (FZD4KO) was used as an FEVR model. (A) Western blot for FZD4 in the retinal tissue of FZD4 knockout mice, compared to wildtype controls (WT). (B–C) Electroretinogram (ERG) assessments at 16 weeks, shown by representative chars (B) and by quantification (C). Blue circle: b-wave; green circle: a-wave. (D) Retinal vascular density by CD31 + area quantification. N = 5. ∗p < 0.05. NS: no significant.

    Journal: Biochemistry and Biophysics Reports

    Article Title: Comparative analysis of activation of macrophages/microglia in diabetic retinopathy and Familial Exudative Vitreoretinopathy

    doi: 10.1016/j.bbrep.2025.102396

    Figure Lengend Snippet: FZD4 knockout mice develop FEVR features. FZD4 knockout mice (FZD4KO) was used as an FEVR model. (A) Western blot for FZD4 in the retinal tissue of FZD4 knockout mice, compared to wildtype controls (WT). (B–C) Electroretinogram (ERG) assessments at 16 weeks, shown by representative chars (B) and by quantification (C). Blue circle: b-wave; green circle: a-wave. (D) Retinal vascular density by CD31 + area quantification. N = 5. ∗p < 0.05. NS: no significant.

    Article Snippet: Knockout of FZD4 in these mice were validated by Western blot for FZD4 in the retinal tissue, using monoclonal antibody against mouse FZD4 (MAB195–050, R&D Systems).

    Techniques: Knock-Out, Western Blot

    Rnf43 regulates the level of TJ through Fzd4. A) Immunofluorescent staining for Zo‐1 with bEnd.3 lacking Fzd4, Fzd6 or Fzd10. Scale bar, 50 µm. B) WB detection for TJ proteins and β‐catenin with bEnd.3 lacking Fzd4, Fzd6 or Fzd10. C) Statistic analysis of relative intensity of TJ proteins and β‐catenin in WB detection, n=4. D) Co‐IP detection of Rnf43 and Fzd4. E) Co‐location detection of Rnf43 and Fzd4. Scale bar, 10 µm. F) WB detection for Fzd4 after Rnf43 knockdown. G) Statistic analysis of relative intensity of Fzd4 in WB detection, n=4. H) Detection of Fzd4 after Rnf43 knockdown. Scale bar, 10 µm. I) Statistic analysis of relative signal intensity of Fzd4, n=4. Data were presented as mean ± SEM, one‐way ANOVA, Two‐tailed Student's t ‐test, ns, no significant difference, * p < 0.05, ** p < 0.01, *** p < 0.001.

    Journal: Advanced Science

    Article Title: Brd4 BD1 Domain Antagonism of MS436 Preserves Blood‐Brain Barrier Integrity via Rnf43/β‐Catenin Signaling Pathway

    doi: 10.1002/advs.202515584

    Figure Lengend Snippet: Rnf43 regulates the level of TJ through Fzd4. A) Immunofluorescent staining for Zo‐1 with bEnd.3 lacking Fzd4, Fzd6 or Fzd10. Scale bar, 50 µm. B) WB detection for TJ proteins and β‐catenin with bEnd.3 lacking Fzd4, Fzd6 or Fzd10. C) Statistic analysis of relative intensity of TJ proteins and β‐catenin in WB detection, n=4. D) Co‐IP detection of Rnf43 and Fzd4. E) Co‐location detection of Rnf43 and Fzd4. Scale bar, 10 µm. F) WB detection for Fzd4 after Rnf43 knockdown. G) Statistic analysis of relative intensity of Fzd4 in WB detection, n=4. H) Detection of Fzd4 after Rnf43 knockdown. Scale bar, 10 µm. I) Statistic analysis of relative signal intensity of Fzd4, n=4. Data were presented as mean ± SEM, one‐way ANOVA, Two‐tailed Student's t ‐test, ns, no significant difference, * p < 0.05, ** p < 0.01, *** p < 0.001.

    Article Snippet: The following primary antibodies and dilutions were used for immunostaining and western blotting: Brd4 (Abcam ab128874, Rabbit, 1:200); Brd4 (Invitrogen A301‐985A‐T, Rabbit, 1:500); Zo‐1 (Invitrogen 40‐2200, Rabbit, 1:500); Zo‐1 (Proteintech 21773‐1‐AP, Rabbit, 1:500); Claudin 5 (Invitrogen 352 500, Mouse, 1:500); Claudin 5 (Abclonal A10207, Rabbit, 1:500); GFAP (Sigma, G6171, Mouse, 1:1000); β‐catenin (Cell Signaling Technology, 8480s, Rabbit, 1:500); Fzd4 (Saribio, K109620P, Rabbit, 1:1 k); biotinylated IsolectinB4 (Vector Laboratories, B‐1205, 1:600); HA (Cell Signaling Technology, 3724s, Rabbit, 1:1 k); Flag (Sigma, F7425, Mouse, 1:2 k); β‐actin (Proteintech, 20536‐1‐AP, Rabbit, 1:10 000); β‐actin (Proteintech; 60008‐1‐Ig Mouse,1:2000); IgG (Bioss, bs‐0295p; Rabbit, 1:1 k).

    Techniques: Staining, Co-Immunoprecipitation Assay, Knockdown, Two Tailed Test

    Brd4 regulates the expression of TJ proteins depends on its BD1 domain. A) Regulation of Claudin 5 by BD domains. Scale bar, 10 µm. B) Immunofluorescent staining for Zo‐1 with bEnd.3 overexpressing BD1 and BD2. Scale bar, 20 µm. C) Statistic analysis of relative intensity of Zo‐1, n=4. D) WB detection for TJ proteins with bEnd.3 overexpressing BD1 or BD2. E) Statistic analysis of relative intensity of TJ proteins in WB detection, n=4. F) Regulation of Claudin 5 by Brd4 mutants. Scale bar, 10 µm. G) Detection of the regulation of Brd4 mutants on the expression of Rnf43. H) Detection of Fzd4 after mutation of Brd4 . Scale bar, 10 µm. Data were presented as mean ± SEM, one‐way ANOVA, Two‐tailed Student's t ‐test, ns, no significant difference, * p < 0.05, ** p < 0.01, *** p < 0.001.

    Journal: Advanced Science

    Article Title: Brd4 BD1 Domain Antagonism of MS436 Preserves Blood‐Brain Barrier Integrity via Rnf43/β‐Catenin Signaling Pathway

    doi: 10.1002/advs.202515584

    Figure Lengend Snippet: Brd4 regulates the expression of TJ proteins depends on its BD1 domain. A) Regulation of Claudin 5 by BD domains. Scale bar, 10 µm. B) Immunofluorescent staining for Zo‐1 with bEnd.3 overexpressing BD1 and BD2. Scale bar, 20 µm. C) Statistic analysis of relative intensity of Zo‐1, n=4. D) WB detection for TJ proteins with bEnd.3 overexpressing BD1 or BD2. E) Statistic analysis of relative intensity of TJ proteins in WB detection, n=4. F) Regulation of Claudin 5 by Brd4 mutants. Scale bar, 10 µm. G) Detection of the regulation of Brd4 mutants on the expression of Rnf43. H) Detection of Fzd4 after mutation of Brd4 . Scale bar, 10 µm. Data were presented as mean ± SEM, one‐way ANOVA, Two‐tailed Student's t ‐test, ns, no significant difference, * p < 0.05, ** p < 0.01, *** p < 0.001.

    Article Snippet: The following primary antibodies and dilutions were used for immunostaining and western blotting: Brd4 (Abcam ab128874, Rabbit, 1:200); Brd4 (Invitrogen A301‐985A‐T, Rabbit, 1:500); Zo‐1 (Invitrogen 40‐2200, Rabbit, 1:500); Zo‐1 (Proteintech 21773‐1‐AP, Rabbit, 1:500); Claudin 5 (Invitrogen 352 500, Mouse, 1:500); Claudin 5 (Abclonal A10207, Rabbit, 1:500); GFAP (Sigma, G6171, Mouse, 1:1000); β‐catenin (Cell Signaling Technology, 8480s, Rabbit, 1:500); Fzd4 (Saribio, K109620P, Rabbit, 1:1 k); biotinylated IsolectinB4 (Vector Laboratories, B‐1205, 1:600); HA (Cell Signaling Technology, 3724s, Rabbit, 1:1 k); Flag (Sigma, F7425, Mouse, 1:2 k); β‐actin (Proteintech, 20536‐1‐AP, Rabbit, 1:10 000); β‐actin (Proteintech; 60008‐1‐Ig Mouse,1:2000); IgG (Bioss, bs‐0295p; Rabbit, 1:1 k).

    Techniques: Expressing, Staining, Mutagenesis, Two Tailed Test