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Abcam rat crb3

EPEC induces internalization of Crb polarity proteins <t>Crb3</t> and Pals1, but not Patj in intestinal epithelial cells. SKCO-15 cells were plated on Transwells and infected on the apical side, or not (UI), with EPEC for 2h. Immunolocalization of endogenous Crb3/Pals1/Patj and E-cad was performed. (A and B) Representative confocal images and quantification of the fluorescence intensity are shown. Arrows indicate the loss of membrane-associated Crb3 and Pals1 induced by EPEC. Hoechst was used to mark the nuclei (blue) in all the images. Scale bar, 10 μm. Data represent the mean ± SEM (n=3); ***P < 0.001 values were calculated using ANOVA Tukey’s Multiple Comparison Test.

Rat Crb3, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 265 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/rat+crb3/Anti-IL-1F10+antibody+-+C-terminal/pmc05645036-339-7-9
Average 99 stars, based on 265 article reviews

rat crb3 - by Bioz Stars, 2026-09

99/100 stars

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1) Product Images from "EPEC effector EspF promotes Crumbs3 endocytosis and disrupts epithelial cell polarity"

Article Title: EPEC effector EspF promotes Crumbs3 endocytosis and disrupts epithelial cell polarity

Journal: Cellular microbiology

doi: 10.1111/cmi.12757

EPEC induces internalization of Crb polarity proteins Crb3 and Pals1, but not Patj in intestinal epithelial cells. SKCO-15 cells were plated on Transwells and infected on the apical side, or not (UI), with EPEC for 2h. Immunolocalization of endogenous Crb3/Pals1/Patj and E-cad was performed. (A and B) Representative confocal images and quantification of the fluorescence intensity are shown. Arrows indicate the loss of membrane-associated Crb3 and Pals1 induced by EPEC. Hoechst was used to mark the nuclei (blue) in all the images. Scale bar, 10 μm. Data represent the mean ± SEM (n=3); ***P < 0.001 values were calculated using ANOVA Tukey’s Multiple Comparison Test.

Figure Legend Snippet: EPEC induces internalization of Crb polarity proteins Crb3 and Pals1, but not Patj in intestinal epithelial cells. SKCO-15 cells were plated on Transwells and infected on the apical side, or not (UI), with EPEC for 2h. Immunolocalization of endogenous Crb3/Pals1/Patj and E-cad was performed. (A and B) Representative confocal images and quantification of the fluorescence intensity are shown. Arrows indicate the loss of membrane-associated Crb3 and Pals1 induced by EPEC. Hoechst was used to mark the nuclei (blue) in all the images. Scale bar, 10 μm. Data represent the mean ± SEM (n=3); ***P < 0.001 values were calculated using ANOVA Tukey’s Multiple Comparison Test.

Techniques Used: Infection, Fluorescence

EPEC and C. rodentium redistribute Crb complex proteins from the plasma membrane to the cytoplasm of murine colonocytes. Mice were infected with EPEC or C. rodentium strain by oral gavage, sacrificed on day 3 or day 10 post-infection respectively; intestinal tissues were processed for immunofluorescence. (A–D) Representative confocal images of Crb3/Pals1/Patj and quantification of the fluorescence intensity of the colonic tissues are shown. Scale bar, 40 μm. Data represent the mean ± SEM (n=3); ***P < 0.001 values were calculated using ANOVA Tukey’s Multiple Comparison Test.

Figure Legend Snippet: EPEC and C. rodentium redistribute Crb complex proteins from the plasma membrane to the cytoplasm of murine colonocytes. Mice were infected with EPEC or C. rodentium strain by oral gavage, sacrificed on day 3 or day 10 post-infection respectively; intestinal tissues were processed for immunofluorescence. (A–D) Representative confocal images of Crb3/Pals1/Patj and quantification of the fluorescence intensity of the colonic tissues are shown. Scale bar, 40 μm. Data represent the mean ± SEM (n=3); ***P < 0.001 values were calculated using ANOVA Tukey’s Multiple Comparison Test.

Techniques Used: Infection, Immunofluorescence, Fluorescence

Deletion of espF protects against Crb3 mislocalization while deletion of either map or espF attenuates the internalization of Pals1. SKCO-15 cells were plated on Transwells and infected apically with wild-type EPEC, ΔespF, Δmap, or complemented strains (ΔespF/pespF and Δmap/pmap) for 2h. Cells were immunostained for Crb3/Pals1/Patj and E-cad was used to label the lateral membrane. (A–F) Representative confocal images and quantification of the fluorescence intensity of membrane, cytoplasm, and total protein are shown. Arrows show the absence of membrane-associated Crb3 and Pals1 in infected monolayers. Scale bar, 10 μm. Data represent the mean ± SEM (n=3); ***P < 0.001 values were calculated using ANOVA Tukey’s Multiple Comparison Test.

Figure Legend Snippet: Deletion of espF protects against Crb3 mislocalization while deletion of either map or espF attenuates the internalization of Pals1. SKCO-15 cells were plated on Transwells and infected apically with wild-type EPEC, ΔespF, Δmap, or complemented strains (ΔespF/pespF and Δmap/pmap) for 2h. Cells were immunostained for Crb3/Pals1/Patj and E-cad was used to label the lateral membrane. (A–F) Representative confocal images and quantification of the fluorescence intensity of membrane, cytoplasm, and total protein are shown. Arrows show the absence of membrane-associated Crb3 and Pals1 in infected monolayers. Scale bar, 10 μm. Data represent the mean ± SEM (n=3); ***P < 0.001 values were calculated using ANOVA Tukey’s Multiple Comparison Test.

Techniques Used: Infection, Fluorescence

Ectopic expression of EspF, but not Map, displaces Crb3 from the membrane. A) espF-HA or map-HA was cloned into the doxycycline-inducible pRetroX-Tight-Pur vector and transfected into SKCO-15 cells. Tet-On SKCO-15 cells were plated in absence of doxycycline (−dox) and transgene expression was induced with doxycycline (+dox) for 3 days. B) SKCO-15 cells were transiently transfected or not (control), with GFP-vector or GFP-EspF. Ectopic expression of EspF or Map was evaluated by immunodetection of HA (Green) or GFP (A and B, respectively). The impact of EspF or Map on localization of Crb3 and Patj was evaluated. Scale bar, 10 μm.

Figure Legend Snippet: Ectopic expression of EspF, but not Map, displaces Crb3 from the membrane. A) espF-HA or map-HA was cloned into the doxycycline-inducible pRetroX-Tight-Pur vector and transfected into SKCO-15 cells. Tet-On SKCO-15 cells were plated in absence of doxycycline (−dox) and transgene expression was induced with doxycycline (+dox) for 3 days. B) SKCO-15 cells were transiently transfected or not (control), with GFP-vector or GFP-EspF. Ectopic expression of EspF or Map was evaluated by immunodetection of HA (Green) or GFP (A and B, respectively). The impact of EspF or Map on localization of Crb3 and Patj was evaluated. Scale bar, 10 μm.

Techniques Used: Expressing, Clone Assay, Plasmid Preparation, Transfection, Immunodetection

Depletion of espF or treatment with dynasore inhibits EPEC-induced cytoplasmic internalization of Crb3. (A–B) SKCO-15 cells were infected with wild-type EPEC, ΔespF or complemented strain (ΔespF/pespF) for 4h and processed for immunodetection of Crb3 and Rab5. (A and B) Representative confocal images and quantification of the area of the co-localization vesicles Crb3/Rab5 are shown. Data represent the mean ± SEM (n=3); ***P < 0.001 values were calculated using ANOVA Tukey’s Multiple Comparison Test. C) SKCO-15 cells were plated in Transwells and treated with DMSO or Dyn, then cells were infected or not (UI) with EPEC for 2h. Samples were processed for immunodetection of Crb3. Scale bar, 10 μm.

Figure Legend Snippet: Depletion of espF or treatment with dynasore inhibits EPEC-induced cytoplasmic internalization of Crb3. (A–B) SKCO-15 cells were infected with wild-type EPEC, ΔespF or complemented strain (ΔespF/pespF) for 4h and processed for immunodetection of Crb3 and Rab5. (A and B) Representative confocal images and quantification of the area of the co-localization vesicles Crb3/Rab5 are shown. Data represent the mean ± SEM (n=3); ***P < 0.001 values were calculated using ANOVA Tukey’s Multiple Comparison Test. C) SKCO-15 cells were plated in Transwells and treated with DMSO or Dyn, then cells were infected or not (UI) with EPEC for 2h. Samples were processed for immunodetection of Crb3. Scale bar, 10 μm.

Techniques Used: Infection, Immunodetection

The interaction of EspF with SNX9 is crucial for Crb3 endocytosis. SKCO-15 cells were infected with wild-type EPEC, ΔespF, or ΔespF complemented to express specific site-directed EspF mutations (ΔespF/pSer47A, ΔespF/pSer47/50A and ΔespF/pespFD3) for 2h. (A and B) Representative images of Crb3 localization and quantification of the fluorescence intensity are shown. Only infection with ΔespF and ΔespF/pespF-D3, which cannot bind SNX9, preserved Crb3 localization. Scale bar, 10 μm. Data represent the mean ± SEM (n=3); ***P < 0.001 values were calculated using ANOVA Tukey’s Multiple Comparison Test.

Figure Legend Snippet: The interaction of EspF with SNX9 is crucial for Crb3 endocytosis. SKCO-15 cells were infected with wild-type EPEC, ΔespF, or ΔespF complemented to express specific site-directed EspF mutations (ΔespF/pSer47A, ΔespF/pSer47/50A and ΔespF/pespFD3) for 2h. (A and B) Representative images of Crb3 localization and quantification of the fluorescence intensity are shown. Only infection with ΔespF and ΔespF/pespF-D3, which cannot bind SNX9, preserved Crb3 localization. Scale bar, 10 μm. Data represent the mean ± SEM (n=3); ***P < 0.001 values were calculated using ANOVA Tukey’s Multiple Comparison Test.

Techniques Used: Infection, Fluorescence

Model depicting the effect of EPEC on apical-basal polarity. Polarized epithelial cells consist of the apical membrane facing the lumen and the basolateral domain contacting the underlying basement membrane. The apical polarity complex Crb (Crb3/Pals1/Patj) localizes at the TJ (Control). i) During EPEC infection, the injected EPEC effectors, including EspF and Map, induce the endocytosis of polarity proteins Crb3 and Pals1 and the basolateral protein Na+/K+ ATPase. ii) Increased endocytosis of Crb3, Pals1 and likely other polarity proteins leads to a loss in the apical-basal polarity, as demonstrated by the redistribution of Na+/K+ ATPase to the apical membrane.

Figure Legend Snippet: Model depicting the effect of EPEC on apical-basal polarity. Polarized epithelial cells consist of the apical membrane facing the lumen and the basolateral domain contacting the underlying basement membrane. The apical polarity complex Crb (Crb3/Pals1/Patj) localizes at the TJ (Control). i) During EPEC infection, the injected EPEC effectors, including EspF and Map, induce the endocytosis of polarity proteins Crb3 and Pals1 and the basolateral protein Na+/K+ ATPase. ii) Increased endocytosis of Crb3, Pals1 and likely other polarity proteins leads to a loss in the apical-basal polarity, as demonstrated by the redistribution of Na+/K+ ATPase to the apical membrane.

Techniques Used: Infection, Injection

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Transwell system with serous-acinar cells grown on a Transwell insert that separates the apical (saliva) and basolateral (plasma) compartments (A). Confocal microscopy of SGECs grown on Nunc Tech II chamber slides 6 days post-seeding (B). DAPI (nuclear stain) shown in blue (top left) and Crumbs apical marker of polarity (CRB3) in green (top center). Overlay of nuclear stain and CRB3 (top right), 3-D rendering (bottom left), cross-sectional x–z axis (bottom center), and cross-sectional y–z axis (bottom right) identify CRB3 localization on the apical surface of SGECs. Scale bar = 10 µm (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article).

Journal: Toxicology

Article Title: Evaluation of non-invasive biomonitoring of 2,4-Dichlorophenoxyacetic acid (2,4-D) in saliva

doi: 10.1016/j.tox.2018.08.003

Figure Lengend Snippet: Transwell system with serous-acinar cells grown on a Transwell insert that separates the apical (saliva) and basolateral (plasma) compartments (A). Confocal microscopy of SGECs grown on Nunc Tech II chamber slides 6 days post-seeding (B). DAPI (nuclear stain) shown in blue (top left) and Crumbs apical marker of polarity (CRB3) in green (top center). Overlay of nuclear stain and CRB3 (top right), 3-D rendering (bottom left), cross-sectional x–z axis (bottom center), and cross-sectional y–z axis (bottom right) identify CRB3 localization on the apical surface of SGECs. Scale bar = 10 µm (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article).

Article Snippet: Anti-CRB3 rat primary monoclonal antibody was purchased from MyBioSource (Cat # MBS588104) and goat anti-rat IgG secondary antibody-Alexa Fluor 488 conjugate was purchased from Invitrogen (Cat # {"type":"entrez-nucleotide","attrs":{"text":"A11006","term_id":"492389","term_text":"A11006"}} A11006 ).

Techniques: Clinical Proteomics, Confocal Microscopy, Staining, Marker

Journal: Cellular microbiology

Article Title: EPEC effector EspF promotes Crumbs3 endocytosis and disrupts epithelial cell polarity

doi: 10.1111/cmi.12757

Figure Lengend Snippet: EPEC induces internalization of Crb polarity proteins Crb3 and Pals1, but not Patj in intestinal epithelial cells. SKCO-15 cells were plated on Transwells and infected on the apical side, or not (UI), with EPEC for 2h. Immunolocalization of endogenous Crb3/Pals1/Patj and E-cad was performed. (A and B) Representative confocal images and quantification of the fluorescence intensity are shown. Arrows indicate the loss of membrane-associated Crb3 and Pals1 induced by EPEC. Hoechst was used to mark the nuclei (blue) in all the images. Scale bar, 10 μm. Data represent the mean ± SEM (n=3); ***P < 0.001 values were calculated using ANOVA Tukey’s Multiple Comparison Test.

Article Snippet: Antibodies used include (species-antigen): rat-Crb3 (ab180835, Abcam), rabbit-Patj (ab102113, Abcam), rabbit-Pals1 (07-708, Millipore), rabbit-ZO-1 (61-7300, Zymed), rabbit-EspF (Hecht Lab), rabbit-cleaved caspase-3 (9661S, Cell Signaling), rabbit-actin (A2066, Sigma), rabbit-HA (C29F4, Cell Signaling), rabbit-GFP ( {"type":"entrez-nucleotide","attrs":{"text":"A11122","term_id":"490966","term_text":"A11122"}} A11122 , Life Technologies), mouse-Rab5 (sc-46692, Santa Cruz), mouse-E-cad (610181, BD), mouse-occludin (33-1500, ThermoFisher Scientific), mouse-Na + /K + ATPase (610992, BD), Phalloidin Alexa Fluor 568 (Life Technologies).

Techniques: Infection, Fluorescence, Membrane, Comparison

Journal: Cellular microbiology

Article Title: EPEC effector EspF promotes Crumbs3 endocytosis and disrupts epithelial cell polarity

doi: 10.1111/cmi.12757

Figure Lengend Snippet: EPEC and C. rodentium redistribute Crb complex proteins from the plasma membrane to the cytoplasm of murine colonocytes. Mice were infected with EPEC or C. rodentium strain by oral gavage, sacrificed on day 3 or day 10 post-infection respectively; intestinal tissues were processed for immunofluorescence. (A–D) Representative confocal images of Crb3/Pals1/Patj and quantification of the fluorescence intensity of the colonic tissues are shown. Scale bar, 40 μm. Data represent the mean ± SEM (n=3); ***P < 0.001 values were calculated using ANOVA Tukey’s Multiple Comparison Test.

Techniques: Clinical Proteomics, Membrane, Infection, Immunofluorescence, Fluorescence, Comparison

Journal: Cellular microbiology

Article Title: EPEC effector EspF promotes Crumbs3 endocytosis and disrupts epithelial cell polarity

doi: 10.1111/cmi.12757

Figure Lengend Snippet: Deletion of espF protects against Crb3 mislocalization while deletion of either map or espF attenuates the internalization of Pals1. SKCO-15 cells were plated on Transwells and infected apically with wild-type EPEC, ΔespF, Δmap, or complemented strains (ΔespF/pespF and Δmap/pmap) for 2h. Cells were immunostained for Crb3/Pals1/Patj and E-cad was used to label the lateral membrane. (A–F) Representative confocal images and quantification of the fluorescence intensity of membrane, cytoplasm, and total protein are shown. Arrows show the absence of membrane-associated Crb3 and Pals1 in infected monolayers. Scale bar, 10 μm. Data represent the mean ± SEM (n=3); ***P < 0.001 values were calculated using ANOVA Tukey’s Multiple Comparison Test.

Techniques: Infection, Membrane, Fluorescence, Comparison

Journal: Cellular microbiology

Article Title: EPEC effector EspF promotes Crumbs3 endocytosis and disrupts epithelial cell polarity

doi: 10.1111/cmi.12757

Figure Lengend Snippet: Ectopic expression of EspF, but not Map, displaces Crb3 from the membrane. A) espF-HA or map-HA was cloned into the doxycycline-inducible pRetroX-Tight-Pur vector and transfected into SKCO-15 cells. Tet-On SKCO-15 cells were plated in absence of doxycycline (−dox) and transgene expression was induced with doxycycline (+dox) for 3 days. B) SKCO-15 cells were transiently transfected or not (control), with GFP-vector or GFP-EspF. Ectopic expression of EspF or Map was evaluated by immunodetection of HA (Green) or GFP (A and B, respectively). The impact of EspF or Map on localization of Crb3 and Patj was evaluated. Scale bar, 10 μm.

Techniques: Expressing, Membrane, Clone Assay, Plasmid Preparation, Transfection, Control, Immunodetection

Journal: Cellular microbiology

Article Title: EPEC effector EspF promotes Crumbs3 endocytosis and disrupts epithelial cell polarity

doi: 10.1111/cmi.12757

Figure Lengend Snippet: Depletion of espF or treatment with dynasore inhibits EPEC-induced cytoplasmic internalization of Crb3. (A–B) SKCO-15 cells were infected with wild-type EPEC, ΔespF or complemented strain (ΔespF/pespF) for 4h and processed for immunodetection of Crb3 and Rab5. (A and B) Representative confocal images and quantification of the area of the co-localization vesicles Crb3/Rab5 are shown. Data represent the mean ± SEM (n=3); ***P < 0.001 values were calculated using ANOVA Tukey’s Multiple Comparison Test. C) SKCO-15 cells were plated in Transwells and treated with DMSO or Dyn, then cells were infected or not (UI) with EPEC for 2h. Samples were processed for immunodetection of Crb3. Scale bar, 10 μm.

Techniques: Infection, Immunodetection, Comparison

Journal: Cellular microbiology

Article Title: EPEC effector EspF promotes Crumbs3 endocytosis and disrupts epithelial cell polarity

doi: 10.1111/cmi.12757

Figure Lengend Snippet: The interaction of EspF with SNX9 is crucial for Crb3 endocytosis. SKCO-15 cells were infected with wild-type EPEC, ΔespF, or ΔespF complemented to express specific site-directed EspF mutations (ΔespF/pSer47A, ΔespF/pSer47/50A and ΔespF/pespFD3) for 2h. (A and B) Representative images of Crb3 localization and quantification of the fluorescence intensity are shown. Only infection with ΔespF and ΔespF/pespF-D3, which cannot bind SNX9, preserved Crb3 localization. Scale bar, 10 μm. Data represent the mean ± SEM (n=3); ***P < 0.001 values were calculated using ANOVA Tukey’s Multiple Comparison Test.

Techniques: Infection, Fluorescence, Comparison

Journal: Cellular microbiology

Article Title: EPEC effector EspF promotes Crumbs3 endocytosis and disrupts epithelial cell polarity

doi: 10.1111/cmi.12757

Figure Lengend Snippet: Model depicting the effect of EPEC on apical-basal polarity. Polarized epithelial cells consist of the apical membrane facing the lumen and the basolateral domain contacting the underlying basement membrane. The apical polarity complex Crb (Crb3/Pals1/Patj) localizes at the TJ (Control). i) During EPEC infection, the injected EPEC effectors, including EspF and Map, induce the endocytosis of polarity proteins Crb3 and Pals1 and the basolateral protein Na+/K+ ATPase. ii) Increased endocytosis of Crb3, Pals1 and likely other polarity proteins leads to a loss in the apical-basal polarity, as demonstrated by the redistribution of Na+/K+ ATPase to the apical membrane.

Techniques: Membrane, Control, Infection, Injection

Journal: Cellular microbiology

Article Title: EPEC effector EspF promotes Crumbs3 endocytosis and disrupts epithelial cell polarity

doi: 10.1111/cmi.12757

Article Snippet: Antibodies and Reagents Antibodies used include (species-antigen): rat-Crb3 (ab180835, Abcam), rabbit-Patj (ab102113, Abcam), rabbit-Pals1 (07-708, Millipore), rabbit-ZO-1 (61-7300, Zymed), rabbit-EspF (Hecht Lab), rabbit-cleaved caspase-3 (9661S, Cell Signaling), rabbit-actin (A2066, Sigma), rabbit-HA (C29F4, Cell Signaling), rabbit-GFP ( {"type":"entrez-nucleotide","attrs":{"text":"A11122","term_id":"490966","term_text":"A11122"}} A11122 , Life Technologies), mouse-Rab5 (sc-46692, Santa Cruz), mouse-E-cad (610181, BD), mouse-occludin (33-1500, ThermoFisher Scientific), mouse-Na + /K + ATPase (610992, BD), Phalloidin Alexa Fluor 568 (Life Technologies).

Techniques: Infection, Fluorescence

Journal: Cellular microbiology

Article Title: EPEC effector EspF promotes Crumbs3 endocytosis and disrupts epithelial cell polarity

doi: 10.1111/cmi.12757

Techniques: Infection, Immunofluorescence, Fluorescence

Journal: Cellular microbiology

Article Title: EPEC effector EspF promotes Crumbs3 endocytosis and disrupts epithelial cell polarity

doi: 10.1111/cmi.12757

Techniques: Infection, Fluorescence

Journal: Cellular microbiology

Article Title: EPEC effector EspF promotes Crumbs3 endocytosis and disrupts epithelial cell polarity

doi: 10.1111/cmi.12757

Techniques: Expressing, Clone Assay, Plasmid Preparation, Transfection, Immunodetection

Journal: Cellular microbiology

Article Title: EPEC effector EspF promotes Crumbs3 endocytosis and disrupts epithelial cell polarity

doi: 10.1111/cmi.12757

Techniques: Infection, Immunodetection

Journal: Cellular microbiology

Article Title: EPEC effector EspF promotes Crumbs3 endocytosis and disrupts epithelial cell polarity

doi: 10.1111/cmi.12757

Techniques: Infection, Fluorescence

Journal: Cellular microbiology

Article Title: EPEC effector EspF promotes Crumbs3 endocytosis and disrupts epithelial cell polarity

doi: 10.1111/cmi.12757

Techniques: Infection, Injection

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