rat anti il 38 il 1f10 Search Results


rat crb3  (Danaher Inc)
99
Danaher Inc 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 Danaher Inc, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/rat+anti+il+38+il+1f10/Anti-IL-1F10+antibody+-+C-terminal/pmc05645036-378-4-6
Average 99 stars, based on 1 article reviews
rat crb3 - by Bioz Stars, 2026-09
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pdcd1 antibody / pd-1 / pd1  (NSJ Bioreagents)
99
NSJ Bioreagents pdcd1 antibody / pd-1 / pd1
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.
Pdcd1 Antibody / Pd 1 / Pd1, supplied by NSJ Bioreagents, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/rat+anti+il+38+il+1f10/PDCD1+Antibody+%2F+PD-1+%2F+PD1/custom%40v8347%4010%2E1080%2F2162402x%2E2021%2E1933808
Average 99 stars, based on 1 article reviews
pdcd1 antibody / pd-1 / pd1 - by Bioz Stars, 2026-09
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anti rat endothelial cell antigen  (Bio-Rad)
93
Bio-Rad anti rat endothelial cell antigen
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.
Anti Rat Endothelial Cell Antigen, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/rat+anti+il+38+il+1f10/Rat+anti+Human+Endothelial+Cells+(Fully+Differentiated)/pm24603294-169-12-17
Average 93 stars, based on 1 article reviews
anti rat endothelial cell antigen - by Bioz Stars, 2026-09
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il 10  (Proteintech)
93
Proteintech il 10
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.
Il 10, supplied by Proteintech, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/rat+anti+il+38+il+1f10/IL-1F10+Antibody/pm38380546-376-103-111
Average 93 stars, based on 1 article reviews
il 10 - by Bioz Stars, 2026-09
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anti-mouse nk marker, nk1.1  (Becton Dickinson)
90
Becton Dickinson anti-mouse nk marker, nk1.1
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.
Anti Mouse Nk Marker, Nk1.1, supplied by Becton Dickinson, 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/rat+anti+il+38+il+1f10/propidium+iodide/pmc02967271-86-56-57
Average 90 stars, based on 1 article reviews
anti-mouse nk marker, nk1.1 - by Bioz Stars, 2026-09
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murine anti-il-18bp  (Abnova)
90
Abnova murine anti-il-18bp
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.
Murine Anti Il 18bp, supplied by Abnova, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 90 stars, based on 1 article reviews
murine anti-il-18bp - by Bioz Stars, 2026-09
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anti-il-18  (MBL International)
90
MBL International anti-il-18
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.
Anti Il 18, supplied by MBL International, 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/rat+anti+il+38+il+1f10/il+18+cytokine/pmc04557384-90-7-28
Average 90 stars, based on 1 article reviews
anti-il-18 - by Bioz Stars, 2026-09
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anti pd 1  (Bio X Cell)
98
Bio X Cell anti pd 1
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.
Anti Pd 1, supplied by Bio X Cell, used in various techniques. Bioz Stars score: 98/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/rat+anti+il+38+il+1f10/InVivoMAb+rat+IgG2a+isotype+control%2C+anti-trinitrophenol/10__1080_slash_2162402x__2021__1933808-54-14-17
Average 98 stars, based on 1 article reviews
anti pd 1 - by Bioz Stars, 2026-09
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anti β actin  (Cell Signaling Technology Inc)
94
Cell Signaling Technology Inc anti β actin
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.
Anti β Actin, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/rat+anti+il+38+il+1f10/beta-Actin+Rabbit+mAb/pmc10991933-171-14-20
Average 94 stars, based on 1 article reviews
anti β actin - by Bioz Stars, 2026-09
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rabbit anti il 1racp polyclonal  (Sino Biological)
90
Sino Biological rabbit anti il 1racp polyclonal
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.
Rabbit Anti Il 1racp Polyclonal, supplied by Sino Biological, 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/rat+anti+il+38+il+1f10/IL-1RAcP+%2F+IL-1R3+Antibody%2C+Rabbit+PAb/pmc04557384-164-17-20
Average 90 stars, based on 1 article reviews
rabbit anti il 1racp polyclonal - by Bioz Stars, 2026-09
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rabbit anti vdac antibody  (Cell Signaling Technology Inc)
96
Cell Signaling Technology Inc rabbit anti vdac antibody
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.
Rabbit Anti Vdac Antibody, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/rat+anti+il+38+il+1f10/VDAC+Antibody/pmc11463958-107-21-24
Average 96 stars, based on 1 article reviews
rabbit anti vdac antibody - by Bioz Stars, 2026-09
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rabbit anti parp antibody  (Cell Signaling Technology Inc)
97
Cell Signaling Technology Inc rabbit anti parp antibody
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.
Rabbit Anti Parp Antibody, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/rat+anti+il+38+il+1f10/PARP+Rabbit+mAb/pmc11463958-107-31-34
Average 97 stars, based on 1 article reviews
rabbit anti parp antibody - by Bioz Stars, 2026-09
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Image Search Results


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.

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

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.

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.

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

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.

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.

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, Membrane, Fluorescence, Comparison

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.

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.

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: Expressing, Membrane, Clone Assay, Plasmid Preparation, Transfection, Control, 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.

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.

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, Immunodetection, Comparison

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.

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.

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, Comparison

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.

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.

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: Membrane, Control, Infection, Injection