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rabbit anti-human alk1 antibody  (Millipore)

 
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    Millipore rabbit anti-human alk1 antibody
    Twenty-Seven Predicted miR-31-5p Target Genes That Were Down-Regulated Significantly in Colonic Mucosa of High-miR-31-5p CD Patients Compared With NIBD Controls
    Rabbit Anti Human Alk1 Antibody, supplied by Millipore, 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/result/rabbit anti-human alk1 antibody/product/Millipore
    Average 90 stars, based on 1 article reviews
    rabbit anti-human alk1 antibody - by Bioz Stars, 2026-02
    90/100 stars

    Images

    1) Product Images from "Decreased Colonic Activin Receptor-Like Kinase 1 Disrupts Epithelial Barrier Integrity in Patients With Crohn’s Disease"

    Article Title: Decreased Colonic Activin Receptor-Like Kinase 1 Disrupts Epithelial Barrier Integrity in Patients With Crohn’s Disease

    Journal: Cellular and Molecular Gastroenterology and Hepatology

    doi: 10.1016/j.jcmgh.2020.06.005

    Twenty-Seven Predicted miR-31-5p Target Genes That Were Down-Regulated Significantly in Colonic Mucosa of High-miR-31-5p CD Patients Compared With NIBD Controls
    Figure Legend Snippet: Twenty-Seven Predicted miR-31-5p Target Genes That Were Down-Regulated Significantly in Colonic Mucosa of High-miR-31-5p CD Patients Compared With NIBD Controls

    Techniques Used:

    ALK1 is a target of miR-31-5p in human colonic epithelial cells. ( A ) Correlation of expression in the colonic mucosa of CD patients (N = 10) between miR-31-5p (reads per million miRNAs mapped, RPMMM) and predicted targets of miR-31-5p ( E2F2 , ALK1 , PRKAB1 , and DCBLD2 ; DESeq normalized). ( B ) Association between the expression of miR-31-5p and the expression of ALK1 or E2F2 in isolated colonic epithelial cells (N = 27). Gene expression was quantified by qPCR and samples were split into 3 equally sized groups (N = 9 per group) according to the relative miR-31-5p expression levels. ( C ) Representative blot of ALK1 expression in the colonic tissue of NIBD and CD patients ( left ). Correlation between ALK1 protein expression and miR-31-5p in the colonic mucosa ( right , N = 18). ( D ) ALK1 expression by immunohistochemistry in the colonic mucosa of NIBD controls and CD patients. The values shown at the bottom are the matched miR-31-5p expression level normalized to NIBD. ( E ) 3’UTR reporter assay for ALK1 in the presence or absence of 30 nmol/L miRNA mimics for hsa-miR-31-5p (m31), hsa-miR-122a-5p (m122), or hsa-miR-215-5p (m215) or negative control mimics (NC). N = 6 per group. ( F ) Schematic representation of the miR-31-5p binding sites in the reporter plasmid. ( G ) Site-directed mutagenesis assay with 10 nmol/L of m31 or NC mimics (N = 6 per group). All correlation values were calculated by the Spearman correlation coefficient. Each gene expression was normalized to GAPDH ( ALK1 , E2F2 ) or RNU48 (miR-31-5p). ∗ P < .05. P values were determined by the Kruskal–Wallis test, followed by the Dunn multiple comparison test. Mut, mutation; NC, negative control mimics.
    Figure Legend Snippet: ALK1 is a target of miR-31-5p in human colonic epithelial cells. ( A ) Correlation of expression in the colonic mucosa of CD patients (N = 10) between miR-31-5p (reads per million miRNAs mapped, RPMMM) and predicted targets of miR-31-5p ( E2F2 , ALK1 , PRKAB1 , and DCBLD2 ; DESeq normalized). ( B ) Association between the expression of miR-31-5p and the expression of ALK1 or E2F2 in isolated colonic epithelial cells (N = 27). Gene expression was quantified by qPCR and samples were split into 3 equally sized groups (N = 9 per group) according to the relative miR-31-5p expression levels. ( C ) Representative blot of ALK1 expression in the colonic tissue of NIBD and CD patients ( left ). Correlation between ALK1 protein expression and miR-31-5p in the colonic mucosa ( right , N = 18). ( D ) ALK1 expression by immunohistochemistry in the colonic mucosa of NIBD controls and CD patients. The values shown at the bottom are the matched miR-31-5p expression level normalized to NIBD. ( E ) 3’UTR reporter assay for ALK1 in the presence or absence of 30 nmol/L miRNA mimics for hsa-miR-31-5p (m31), hsa-miR-122a-5p (m122), or hsa-miR-215-5p (m215) or negative control mimics (NC). N = 6 per group. ( F ) Schematic representation of the miR-31-5p binding sites in the reporter plasmid. ( G ) Site-directed mutagenesis assay with 10 nmol/L of m31 or NC mimics (N = 6 per group). All correlation values were calculated by the Spearman correlation coefficient. Each gene expression was normalized to GAPDH ( ALK1 , E2F2 ) or RNU48 (miR-31-5p). ∗ P < .05. P values were determined by the Kruskal–Wallis test, followed by the Dunn multiple comparison test. Mut, mutation; NC, negative control mimics.

    Techniques Used: Expressing, Isolation, Immunohistochemistry, Reporter Assay, Negative Control, Binding Assay, Plasmid Preparation, Mutagenesis

    Decreased ALK1 expression is associated with reduced NOTCH activity and NOTCH target gene expression in the colonic epithelial cells of CD patients. ( A ) Representative blot ( left ) and the difference of JAG1 and NOTCH intracellular domain (NICD) protein expression between NIBD and CD patients ( right ). ( B ) BMP9 concentration in the serum of NIBD controls (N = 17) and CD patients (N = 23). ( C ) NOTCH target gene expression in colonic epithelial cells from CD patients (N = 15) and NIBD controls (N = 12). ( D ) NOTCH target gene expression in NIBD patient-derived colonic epithelial cell monolayers. Expanded cells were cultured in expansion media in the presence or absence of BMP9 and ALK1–Fc chimera protein. N = 6 per group. Each gene expression was normalized to ( C ) GAPDH or ( D ) RPLP0 . ∗ P < .05, ∗∗ P < .01, and ∗∗∗ P < .001. P values were determined by the ( A–C ) Mann–Whitney test or the ( D ) Friedman test followed by the Dunn multiple comparison test.
    Figure Legend Snippet: Decreased ALK1 expression is associated with reduced NOTCH activity and NOTCH target gene expression in the colonic epithelial cells of CD patients. ( A ) Representative blot ( left ) and the difference of JAG1 and NOTCH intracellular domain (NICD) protein expression between NIBD and CD patients ( right ). ( B ) BMP9 concentration in the serum of NIBD controls (N = 17) and CD patients (N = 23). ( C ) NOTCH target gene expression in colonic epithelial cells from CD patients (N = 15) and NIBD controls (N = 12). ( D ) NOTCH target gene expression in NIBD patient-derived colonic epithelial cell monolayers. Expanded cells were cultured in expansion media in the presence or absence of BMP9 and ALK1–Fc chimera protein. N = 6 per group. Each gene expression was normalized to ( C ) GAPDH or ( D ) RPLP0 . ∗ P < .05, ∗∗ P < .01, and ∗∗∗ P < .001. P values were determined by the ( A–C ) Mann–Whitney test or the ( D ) Friedman test followed by the Dunn multiple comparison test.

    Techniques Used: Expressing, Activity Assay, Concentration Assay, Derivative Assay, Cell Culture, MANN-WHITNEY

    Expression of miR-31-5p and ALK1 in primary-cultured colonic epithelial monolayers derived from NIBD controls and CD patients. N = 6 per group. Each gene expression was normalized to RNU48 (miR-31-5p) or GAPDH ( ALK1 ). Statistical significance was determined by the Mann–Whitney test.
    Figure Legend Snippet: Expression of miR-31-5p and ALK1 in primary-cultured colonic epithelial monolayers derived from NIBD controls and CD patients. N = 6 per group. Each gene expression was normalized to RNU48 (miR-31-5p) or GAPDH ( ALK1 ). Statistical significance was determined by the Mann–Whitney test.

    Techniques Used: Expressing, Cell Culture, Derivative Assay, MANN-WHITNEY

    BMP9–ALK1 signaling restricts the stemness of human colonic IECs. ( A ) EdU assay in NIBD patient-derived colonic epithelial cell monolayers (N = 4–8 per group). Expanded cells were cultured in EM in the presence or absence of BMP9 and ALK1–Fc chimera protein. Red, EdU; blue, Hoechst 33342. ( B ) Proliferation- and stemness-related gene expression in NIBD patient-derived colonic epithelial cell monolayers (N = 6 per group). ( C ) Proliferation- and stemness-related gene expression in colonic epithelial cells isolated from CD patients (N = 15) and NIBD controls (N = 12). ( D ) Representative immunohistochemical images of OLFM4 expression in the colonic crypts of NIBD controls ( left ) and CD patients ( right ). The percentage of OLFM4 staining area in colonic crypts was compared between CD patients and NIBD controls (N = 4 per group). Each gene expression was normalized to ( B ) RPLP0 or ( C ) GAPDH . ∗ P < .05, ∗∗ P < .01, and ∗∗∗ P < .001. P values were determined by the ( A ) Kruskal–Wallis test or the ( B ) Friedman test followed by the Dunn multiple comparisons test, or the ( C and D ) Mann–Whitney test.
    Figure Legend Snippet: BMP9–ALK1 signaling restricts the stemness of human colonic IECs. ( A ) EdU assay in NIBD patient-derived colonic epithelial cell monolayers (N = 4–8 per group). Expanded cells were cultured in EM in the presence or absence of BMP9 and ALK1–Fc chimera protein. Red, EdU; blue, Hoechst 33342. ( B ) Proliferation- and stemness-related gene expression in NIBD patient-derived colonic epithelial cell monolayers (N = 6 per group). ( C ) Proliferation- and stemness-related gene expression in colonic epithelial cells isolated from CD patients (N = 15) and NIBD controls (N = 12). ( D ) Representative immunohistochemical images of OLFM4 expression in the colonic crypts of NIBD controls ( left ) and CD patients ( right ). The percentage of OLFM4 staining area in colonic crypts was compared between CD patients and NIBD controls (N = 4 per group). Each gene expression was normalized to ( B ) RPLP0 or ( C ) GAPDH . ∗ P < .05, ∗∗ P < .01, and ∗∗∗ P < .001. P values were determined by the ( A ) Kruskal–Wallis test or the ( B ) Friedman test followed by the Dunn multiple comparisons test, or the ( C and D ) Mann–Whitney test.

    Techniques Used: EdU Assay, Derivative Assay, Cell Culture, Expressing, Isolation, Immunohistochemical staining, Staining, MANN-WHITNEY

    BMP9–ALK1 signaling is associated with epithelial cell differentiation toward colonocytes. ( A ) Lineage-specific gene expression in NIBD patient-derived colonic epithelial cell monolayers. Expanded cells were cultured in expansion media in the presence or absence of BMP9 and ALK1–Fc chimera protein. Each gene expression was normalized to RPLP0 (N = 6 per group). ( B ) CA1 protein expression in NIBD patient-derived colonic epithelial monolayers (N = 4 per group). ( C ) Colonocyte marker expression in colonic epithelial cells isolated from CD patients (N = 15) and NIBD controls (N = 12). Each gene expression was normalized to GAPDH . ( D ) CA1 expression by immunohistochemistry in the colonic mucosa of NIBD controls ( left ) and CD patients ( right ). ( E ) CA1 protein expression in the colonic mucosa of NIBD controls and CD patients (N = 5 per group). ∗ P < .05, ∗∗ P < .01, ∗∗∗ P < .001, and ∗∗∗∗ P < .0001. P values were determined by the ( A and B ) Friedman test followed by the Dunn multiple comparison test, or the ( C and E ) Mann–Whitney test.
    Figure Legend Snippet: BMP9–ALK1 signaling is associated with epithelial cell differentiation toward colonocytes. ( A ) Lineage-specific gene expression in NIBD patient-derived colonic epithelial cell monolayers. Expanded cells were cultured in expansion media in the presence or absence of BMP9 and ALK1–Fc chimera protein. Each gene expression was normalized to RPLP0 (N = 6 per group). ( B ) CA1 protein expression in NIBD patient-derived colonic epithelial monolayers (N = 4 per group). ( C ) Colonocyte marker expression in colonic epithelial cells isolated from CD patients (N = 15) and NIBD controls (N = 12). Each gene expression was normalized to GAPDH . ( D ) CA1 expression by immunohistochemistry in the colonic mucosa of NIBD controls ( left ) and CD patients ( right ). ( E ) CA1 protein expression in the colonic mucosa of NIBD controls and CD patients (N = 5 per group). ∗ P < .05, ∗∗ P < .01, ∗∗∗ P < .001, and ∗∗∗∗ P < .0001. P values were determined by the ( A and B ) Friedman test followed by the Dunn multiple comparison test, or the ( C and E ) Mann–Whitney test.

    Techniques Used: Cell Differentiation, Expressing, Derivative Assay, Cell Culture, Marker, Isolation, Immunohistochemistry, MANN-WHITNEY

    BMP9–ALK1 signaling enhances human colonic IEC barrier integrity. ( A ) Gene expression of junctional proteins in NIBD patient-derived colonic epithelial cell monolayers. Expanded cells were cultured in expansion media in the presence or absence of BMP9 and ALK1–Fc chimera protein. Each gene expression was normalized to RPLP0 (N = 6 per group). ( B ) Gene expression of tight junction proteins in colonic epithelial cells isolated from CD patients (N = 15) and NIBD controls (N = 12). Each gene expression was normalized to GAPDH . ( C ) Epithelial permeability assay in NIBD patient-derived colonic epithelial cells cultured on a collagen scaffold. TEER was measured over time (N = 3 per group). ( D ) Cells were stimulated with BMP9 in EM in the presence or absence of ALK1–Fc chimera protein or cultured in DM on day 4. The changes in TEER between days 4 and 6 are shown as ΔTEER% (N = 4–8 per group). ∗ P < .05, ∗∗ P < .01, and ∗∗∗ P < .001. P values were determined by the ( A and D ) Kruskal–Wallis test followed by the Dunn multiple comparison test, and the ( B ) Mann–Whitney test.
    Figure Legend Snippet: BMP9–ALK1 signaling enhances human colonic IEC barrier integrity. ( A ) Gene expression of junctional proteins in NIBD patient-derived colonic epithelial cell monolayers. Expanded cells were cultured in expansion media in the presence or absence of BMP9 and ALK1–Fc chimera protein. Each gene expression was normalized to RPLP0 (N = 6 per group). ( B ) Gene expression of tight junction proteins in colonic epithelial cells isolated from CD patients (N = 15) and NIBD controls (N = 12). Each gene expression was normalized to GAPDH . ( C ) Epithelial permeability assay in NIBD patient-derived colonic epithelial cells cultured on a collagen scaffold. TEER was measured over time (N = 3 per group). ( D ) Cells were stimulated with BMP9 in EM in the presence or absence of ALK1–Fc chimera protein or cultured in DM on day 4. The changes in TEER between days 4 and 6 are shown as ΔTEER% (N = 4–8 per group). ∗ P < .05, ∗∗ P < .01, and ∗∗∗ P < .001. P values were determined by the ( A and D ) Kruskal–Wallis test followed by the Dunn multiple comparison test, and the ( B ) Mann–Whitney test.

    Techniques Used: Expressing, Derivative Assay, Cell Culture, Isolation, Permeability, MANN-WHITNEY

    Decreased colonic ALK1 is associated with a poor clinical outcome in CD patients. ( A ) ALK1 expression was quantified in colonic biopsy samples obtained from NIBD controls (N = 10) and CD patients (N = 28) by qPCR. ( B ) ALK1 expression in the colonic mucosa of CD patients at the time of surgery and after surgery (N = 5 per group). ( C ) Percentages of CD patients who were diagnosed as CD before and after age 40 years in low-ALK1 (N = 15) and hi-ALK1 (N = 13) CD subsets. ( D ) Kaplan–Meier survival analysis to evaluate the impact of colonic ALK1 expression on endoscopic relapse in patients with CD (N = 12 for low-ALK1 and 9 for hi-ALK1 CD subgroups). ALK1 expression was normalized to GAPDH . ∗∗∗ P < .001. P values were determined by the ( A ) Mann–Whitney test, ( B ) Wilcoxon test, and ( D ) log-rank test.
    Figure Legend Snippet: Decreased colonic ALK1 is associated with a poor clinical outcome in CD patients. ( A ) ALK1 expression was quantified in colonic biopsy samples obtained from NIBD controls (N = 10) and CD patients (N = 28) by qPCR. ( B ) ALK1 expression in the colonic mucosa of CD patients at the time of surgery and after surgery (N = 5 per group). ( C ) Percentages of CD patients who were diagnosed as CD before and after age 40 years in low-ALK1 (N = 15) and hi-ALK1 (N = 13) CD subsets. ( D ) Kaplan–Meier survival analysis to evaluate the impact of colonic ALK1 expression on endoscopic relapse in patients with CD (N = 12 for low-ALK1 and 9 for hi-ALK1 CD subgroups). ALK1 expression was normalized to GAPDH . ∗∗∗ P < .001. P values were determined by the ( A ) Mann–Whitney test, ( B ) Wilcoxon test, and ( D ) log-rank test.

    Techniques Used: Expressing, MANN-WHITNEY

    Clinical Characteristics at the Time of Sample Collection
    Figure Legend Snippet: Clinical Characteristics at the Time of Sample Collection

    Techniques Used: Sampling, Activity Assay

    Correlations Between Explanatory Variables for the Risk of Surgery
    Figure Legend Snippet: Correlations Between Explanatory Variables for the Risk of Surgery

    Techniques Used:

    Binomial Logistic Regression Analysis for Surgery
    Figure Legend Snippet: Binomial Logistic Regression Analysis for Surgery

    Techniques Used:

    Primer Sequences for RT-qPCR
    Figure Legend Snippet: Primer Sequences for RT-qPCR

    Techniques Used:



    Similar Products

    rabbit anti-human alk1 antibody  (Millipore)
    90
    Millipore rabbit anti-human alk1 antibody
    Twenty-Seven Predicted miR-31-5p Target Genes That Were Down-Regulated Significantly in Colonic Mucosa of High-miR-31-5p CD Patients Compared With NIBD Controls
    Rabbit Anti Human Alk1 Antibody, supplied by Millipore, 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/result/rabbit anti-human alk1 antibody/product/Millipore
    Average 90 stars, based on 1 article reviews
    rabbit anti-human alk1 antibody - by Bioz Stars, 2026-02
    90/100 stars
    		  Buy from Supplier

    Image Search Results


    Twenty-Seven Predicted miR-31-5p Target Genes That Were Down-Regulated Significantly in Colonic Mucosa of High-miR-31-5p CD Patients Compared With NIBD Controls

    Journal: Cellular and Molecular Gastroenterology and Hepatology

    Article Title: Decreased Colonic Activin Receptor-Like Kinase 1 Disrupts Epithelial Barrier Integrity in Patients With Crohn’s Disease

    doi: 10.1016/j.jcmgh.2020.06.005

    Figure Lengend Snippet: Twenty-Seven Predicted miR-31-5p Target Genes That Were Down-Regulated Significantly in Colonic Mucosa of High-miR-31-5p CD Patients Compared With NIBD Controls

    Article Snippet: Rabbit anti-human ALK1 antibody, rabbit anti-human CA1 antibody, rabbit anti-human OLFM4 antibody, and mouse/rabbit IgG VisUCyte horseradish-peroxidase polymer antibody were purchased from Sigma-Aldrich (St. Louis, MO) (HPA007041), Novus Biologicals (Littleton, CO) (NBP1-88191), Cell Signaling Technology (Beverly, MA) (14369S), and R&D Systems (VC002-025).

    Techniques:

    ALK1 is a target of miR-31-5p in human colonic epithelial cells. ( A ) Correlation of expression in the colonic mucosa of CD patients (N = 10) between miR-31-5p (reads per million miRNAs mapped, RPMMM) and predicted targets of miR-31-5p ( E2F2 , ALK1 , PRKAB1 , and DCBLD2 ; DESeq normalized). ( B ) Association between the expression of miR-31-5p and the expression of ALK1 or E2F2 in isolated colonic epithelial cells (N = 27). Gene expression was quantified by qPCR and samples were split into 3 equally sized groups (N = 9 per group) according to the relative miR-31-5p expression levels. ( C ) Representative blot of ALK1 expression in the colonic tissue of NIBD and CD patients ( left ). Correlation between ALK1 protein expression and miR-31-5p in the colonic mucosa ( right , N = 18). ( D ) ALK1 expression by immunohistochemistry in the colonic mucosa of NIBD controls and CD patients. The values shown at the bottom are the matched miR-31-5p expression level normalized to NIBD. ( E ) 3’UTR reporter assay for ALK1 in the presence or absence of 30 nmol/L miRNA mimics for hsa-miR-31-5p (m31), hsa-miR-122a-5p (m122), or hsa-miR-215-5p (m215) or negative control mimics (NC). N = 6 per group. ( F ) Schematic representation of the miR-31-5p binding sites in the reporter plasmid. ( G ) Site-directed mutagenesis assay with 10 nmol/L of m31 or NC mimics (N = 6 per group). All correlation values were calculated by the Spearman correlation coefficient. Each gene expression was normalized to GAPDH ( ALK1 , E2F2 ) or RNU48 (miR-31-5p). ∗ P < .05. P values were determined by the Kruskal–Wallis test, followed by the Dunn multiple comparison test. Mut, mutation; NC, negative control mimics.

    Journal: Cellular and Molecular Gastroenterology and Hepatology

    Article Title: Decreased Colonic Activin Receptor-Like Kinase 1 Disrupts Epithelial Barrier Integrity in Patients With Crohn’s Disease

    doi: 10.1016/j.jcmgh.2020.06.005

    Figure Lengend Snippet: ALK1 is a target of miR-31-5p in human colonic epithelial cells. ( A ) Correlation of expression in the colonic mucosa of CD patients (N = 10) between miR-31-5p (reads per million miRNAs mapped, RPMMM) and predicted targets of miR-31-5p ( E2F2 , ALK1 , PRKAB1 , and DCBLD2 ; DESeq normalized). ( B ) Association between the expression of miR-31-5p and the expression of ALK1 or E2F2 in isolated colonic epithelial cells (N = 27). Gene expression was quantified by qPCR and samples were split into 3 equally sized groups (N = 9 per group) according to the relative miR-31-5p expression levels. ( C ) Representative blot of ALK1 expression in the colonic tissue of NIBD and CD patients ( left ). Correlation between ALK1 protein expression and miR-31-5p in the colonic mucosa ( right , N = 18). ( D ) ALK1 expression by immunohistochemistry in the colonic mucosa of NIBD controls and CD patients. The values shown at the bottom are the matched miR-31-5p expression level normalized to NIBD. ( E ) 3’UTR reporter assay for ALK1 in the presence or absence of 30 nmol/L miRNA mimics for hsa-miR-31-5p (m31), hsa-miR-122a-5p (m122), or hsa-miR-215-5p (m215) or negative control mimics (NC). N = 6 per group. ( F ) Schematic representation of the miR-31-5p binding sites in the reporter plasmid. ( G ) Site-directed mutagenesis assay with 10 nmol/L of m31 or NC mimics (N = 6 per group). All correlation values were calculated by the Spearman correlation coefficient. Each gene expression was normalized to GAPDH ( ALK1 , E2F2 ) or RNU48 (miR-31-5p). ∗ P < .05. P values were determined by the Kruskal–Wallis test, followed by the Dunn multiple comparison test. Mut, mutation; NC, negative control mimics.

    Article Snippet: Rabbit anti-human ALK1 antibody, rabbit anti-human CA1 antibody, rabbit anti-human OLFM4 antibody, and mouse/rabbit IgG VisUCyte horseradish-peroxidase polymer antibody were purchased from Sigma-Aldrich (St. Louis, MO) (HPA007041), Novus Biologicals (Littleton, CO) (NBP1-88191), Cell Signaling Technology (Beverly, MA) (14369S), and R&D Systems (VC002-025).

    Techniques: Expressing, Isolation, Immunohistochemistry, Reporter Assay, Negative Control, Binding Assay, Plasmid Preparation, Mutagenesis

    Decreased ALK1 expression is associated with reduced NOTCH activity and NOTCH target gene expression in the colonic epithelial cells of CD patients. ( A ) Representative blot ( left ) and the difference of JAG1 and NOTCH intracellular domain (NICD) protein expression between NIBD and CD patients ( right ). ( B ) BMP9 concentration in the serum of NIBD controls (N = 17) and CD patients (N = 23). ( C ) NOTCH target gene expression in colonic epithelial cells from CD patients (N = 15) and NIBD controls (N = 12). ( D ) NOTCH target gene expression in NIBD patient-derived colonic epithelial cell monolayers. Expanded cells were cultured in expansion media in the presence or absence of BMP9 and ALK1–Fc chimera protein. N = 6 per group. Each gene expression was normalized to ( C ) GAPDH or ( D ) RPLP0 . ∗ P < .05, ∗∗ P < .01, and ∗∗∗ P < .001. P values were determined by the ( A–C ) Mann–Whitney test or the ( D ) Friedman test followed by the Dunn multiple comparison test.

    Journal: Cellular and Molecular Gastroenterology and Hepatology

    Article Title: Decreased Colonic Activin Receptor-Like Kinase 1 Disrupts Epithelial Barrier Integrity in Patients With Crohn’s Disease

    doi: 10.1016/j.jcmgh.2020.06.005

    Figure Lengend Snippet: Decreased ALK1 expression is associated with reduced NOTCH activity and NOTCH target gene expression in the colonic epithelial cells of CD patients. ( A ) Representative blot ( left ) and the difference of JAG1 and NOTCH intracellular domain (NICD) protein expression between NIBD and CD patients ( right ). ( B ) BMP9 concentration in the serum of NIBD controls (N = 17) and CD patients (N = 23). ( C ) NOTCH target gene expression in colonic epithelial cells from CD patients (N = 15) and NIBD controls (N = 12). ( D ) NOTCH target gene expression in NIBD patient-derived colonic epithelial cell monolayers. Expanded cells were cultured in expansion media in the presence or absence of BMP9 and ALK1–Fc chimera protein. N = 6 per group. Each gene expression was normalized to ( C ) GAPDH or ( D ) RPLP0 . ∗ P < .05, ∗∗ P < .01, and ∗∗∗ P < .001. P values were determined by the ( A–C ) Mann–Whitney test or the ( D ) Friedman test followed by the Dunn multiple comparison test.

    Article Snippet: Rabbit anti-human ALK1 antibody, rabbit anti-human CA1 antibody, rabbit anti-human OLFM4 antibody, and mouse/rabbit IgG VisUCyte horseradish-peroxidase polymer antibody were purchased from Sigma-Aldrich (St. Louis, MO) (HPA007041), Novus Biologicals (Littleton, CO) (NBP1-88191), Cell Signaling Technology (Beverly, MA) (14369S), and R&D Systems (VC002-025).

    Techniques: Expressing, Activity Assay, Concentration Assay, Derivative Assay, Cell Culture, MANN-WHITNEY

    Expression of miR-31-5p and ALK1 in primary-cultured colonic epithelial monolayers derived from NIBD controls and CD patients. N = 6 per group. Each gene expression was normalized to RNU48 (miR-31-5p) or GAPDH ( ALK1 ). Statistical significance was determined by the Mann–Whitney test.

    Journal: Cellular and Molecular Gastroenterology and Hepatology

    Article Title: Decreased Colonic Activin Receptor-Like Kinase 1 Disrupts Epithelial Barrier Integrity in Patients With Crohn’s Disease

    doi: 10.1016/j.jcmgh.2020.06.005

    Figure Lengend Snippet: Expression of miR-31-5p and ALK1 in primary-cultured colonic epithelial monolayers derived from NIBD controls and CD patients. N = 6 per group. Each gene expression was normalized to RNU48 (miR-31-5p) or GAPDH ( ALK1 ). Statistical significance was determined by the Mann–Whitney test.

    Article Snippet: Rabbit anti-human ALK1 antibody, rabbit anti-human CA1 antibody, rabbit anti-human OLFM4 antibody, and mouse/rabbit IgG VisUCyte horseradish-peroxidase polymer antibody were purchased from Sigma-Aldrich (St. Louis, MO) (HPA007041), Novus Biologicals (Littleton, CO) (NBP1-88191), Cell Signaling Technology (Beverly, MA) (14369S), and R&D Systems (VC002-025).

    Techniques: Expressing, Cell Culture, Derivative Assay, MANN-WHITNEY

    BMP9–ALK1 signaling restricts the stemness of human colonic IECs. ( A ) EdU assay in NIBD patient-derived colonic epithelial cell monolayers (N = 4–8 per group). Expanded cells were cultured in EM in the presence or absence of BMP9 and ALK1–Fc chimera protein. Red, EdU; blue, Hoechst 33342. ( B ) Proliferation- and stemness-related gene expression in NIBD patient-derived colonic epithelial cell monolayers (N = 6 per group). ( C ) Proliferation- and stemness-related gene expression in colonic epithelial cells isolated from CD patients (N = 15) and NIBD controls (N = 12). ( D ) Representative immunohistochemical images of OLFM4 expression in the colonic crypts of NIBD controls ( left ) and CD patients ( right ). The percentage of OLFM4 staining area in colonic crypts was compared between CD patients and NIBD controls (N = 4 per group). Each gene expression was normalized to ( B ) RPLP0 or ( C ) GAPDH . ∗ P < .05, ∗∗ P < .01, and ∗∗∗ P < .001. P values were determined by the ( A ) Kruskal–Wallis test or the ( B ) Friedman test followed by the Dunn multiple comparisons test, or the ( C and D ) Mann–Whitney test.

    Journal: Cellular and Molecular Gastroenterology and Hepatology

    Article Title: Decreased Colonic Activin Receptor-Like Kinase 1 Disrupts Epithelial Barrier Integrity in Patients With Crohn’s Disease

    doi: 10.1016/j.jcmgh.2020.06.005

    Figure Lengend Snippet: BMP9–ALK1 signaling restricts the stemness of human colonic IECs. ( A ) EdU assay in NIBD patient-derived colonic epithelial cell monolayers (N = 4–8 per group). Expanded cells were cultured in EM in the presence or absence of BMP9 and ALK1–Fc chimera protein. Red, EdU; blue, Hoechst 33342. ( B ) Proliferation- and stemness-related gene expression in NIBD patient-derived colonic epithelial cell monolayers (N = 6 per group). ( C ) Proliferation- and stemness-related gene expression in colonic epithelial cells isolated from CD patients (N = 15) and NIBD controls (N = 12). ( D ) Representative immunohistochemical images of OLFM4 expression in the colonic crypts of NIBD controls ( left ) and CD patients ( right ). The percentage of OLFM4 staining area in colonic crypts was compared between CD patients and NIBD controls (N = 4 per group). Each gene expression was normalized to ( B ) RPLP0 or ( C ) GAPDH . ∗ P < .05, ∗∗ P < .01, and ∗∗∗ P < .001. P values were determined by the ( A ) Kruskal–Wallis test or the ( B ) Friedman test followed by the Dunn multiple comparisons test, or the ( C and D ) Mann–Whitney test.

    Article Snippet: Rabbit anti-human ALK1 antibody, rabbit anti-human CA1 antibody, rabbit anti-human OLFM4 antibody, and mouse/rabbit IgG VisUCyte horseradish-peroxidase polymer antibody were purchased from Sigma-Aldrich (St. Louis, MO) (HPA007041), Novus Biologicals (Littleton, CO) (NBP1-88191), Cell Signaling Technology (Beverly, MA) (14369S), and R&D Systems (VC002-025).

    Techniques: EdU Assay, Derivative Assay, Cell Culture, Expressing, Isolation, Immunohistochemical staining, Staining, MANN-WHITNEY

    BMP9–ALK1 signaling is associated with epithelial cell differentiation toward colonocytes. ( A ) Lineage-specific gene expression in NIBD patient-derived colonic epithelial cell monolayers. Expanded cells were cultured in expansion media in the presence or absence of BMP9 and ALK1–Fc chimera protein. Each gene expression was normalized to RPLP0 (N = 6 per group). ( B ) CA1 protein expression in NIBD patient-derived colonic epithelial monolayers (N = 4 per group). ( C ) Colonocyte marker expression in colonic epithelial cells isolated from CD patients (N = 15) and NIBD controls (N = 12). Each gene expression was normalized to GAPDH . ( D ) CA1 expression by immunohistochemistry in the colonic mucosa of NIBD controls ( left ) and CD patients ( right ). ( E ) CA1 protein expression in the colonic mucosa of NIBD controls and CD patients (N = 5 per group). ∗ P < .05, ∗∗ P < .01, ∗∗∗ P < .001, and ∗∗∗∗ P < .0001. P values were determined by the ( A and B ) Friedman test followed by the Dunn multiple comparison test, or the ( C and E ) Mann–Whitney test.

    Journal: Cellular and Molecular Gastroenterology and Hepatology

    Article Title: Decreased Colonic Activin Receptor-Like Kinase 1 Disrupts Epithelial Barrier Integrity in Patients With Crohn’s Disease

    doi: 10.1016/j.jcmgh.2020.06.005

    Figure Lengend Snippet: BMP9–ALK1 signaling is associated with epithelial cell differentiation toward colonocytes. ( A ) Lineage-specific gene expression in NIBD patient-derived colonic epithelial cell monolayers. Expanded cells were cultured in expansion media in the presence or absence of BMP9 and ALK1–Fc chimera protein. Each gene expression was normalized to RPLP0 (N = 6 per group). ( B ) CA1 protein expression in NIBD patient-derived colonic epithelial monolayers (N = 4 per group). ( C ) Colonocyte marker expression in colonic epithelial cells isolated from CD patients (N = 15) and NIBD controls (N = 12). Each gene expression was normalized to GAPDH . ( D ) CA1 expression by immunohistochemistry in the colonic mucosa of NIBD controls ( left ) and CD patients ( right ). ( E ) CA1 protein expression in the colonic mucosa of NIBD controls and CD patients (N = 5 per group). ∗ P < .05, ∗∗ P < .01, ∗∗∗ P < .001, and ∗∗∗∗ P < .0001. P values were determined by the ( A and B ) Friedman test followed by the Dunn multiple comparison test, or the ( C and E ) Mann–Whitney test.

    Article Snippet: Rabbit anti-human ALK1 antibody, rabbit anti-human CA1 antibody, rabbit anti-human OLFM4 antibody, and mouse/rabbit IgG VisUCyte horseradish-peroxidase polymer antibody were purchased from Sigma-Aldrich (St. Louis, MO) (HPA007041), Novus Biologicals (Littleton, CO) (NBP1-88191), Cell Signaling Technology (Beverly, MA) (14369S), and R&D Systems (VC002-025).

    Techniques: Cell Differentiation, Expressing, Derivative Assay, Cell Culture, Marker, Isolation, Immunohistochemistry, MANN-WHITNEY

    BMP9–ALK1 signaling enhances human colonic IEC barrier integrity. ( A ) Gene expression of junctional proteins in NIBD patient-derived colonic epithelial cell monolayers. Expanded cells were cultured in expansion media in the presence or absence of BMP9 and ALK1–Fc chimera protein. Each gene expression was normalized to RPLP0 (N = 6 per group). ( B ) Gene expression of tight junction proteins in colonic epithelial cells isolated from CD patients (N = 15) and NIBD controls (N = 12). Each gene expression was normalized to GAPDH . ( C ) Epithelial permeability assay in NIBD patient-derived colonic epithelial cells cultured on a collagen scaffold. TEER was measured over time (N = 3 per group). ( D ) Cells were stimulated with BMP9 in EM in the presence or absence of ALK1–Fc chimera protein or cultured in DM on day 4. The changes in TEER between days 4 and 6 are shown as ΔTEER% (N = 4–8 per group). ∗ P < .05, ∗∗ P < .01, and ∗∗∗ P < .001. P values were determined by the ( A and D ) Kruskal–Wallis test followed by the Dunn multiple comparison test, and the ( B ) Mann–Whitney test.

    Journal: Cellular and Molecular Gastroenterology and Hepatology

    Article Title: Decreased Colonic Activin Receptor-Like Kinase 1 Disrupts Epithelial Barrier Integrity in Patients With Crohn’s Disease

    doi: 10.1016/j.jcmgh.2020.06.005

    Figure Lengend Snippet: BMP9–ALK1 signaling enhances human colonic IEC barrier integrity. ( A ) Gene expression of junctional proteins in NIBD patient-derived colonic epithelial cell monolayers. Expanded cells were cultured in expansion media in the presence or absence of BMP9 and ALK1–Fc chimera protein. Each gene expression was normalized to RPLP0 (N = 6 per group). ( B ) Gene expression of tight junction proteins in colonic epithelial cells isolated from CD patients (N = 15) and NIBD controls (N = 12). Each gene expression was normalized to GAPDH . ( C ) Epithelial permeability assay in NIBD patient-derived colonic epithelial cells cultured on a collagen scaffold. TEER was measured over time (N = 3 per group). ( D ) Cells were stimulated with BMP9 in EM in the presence or absence of ALK1–Fc chimera protein or cultured in DM on day 4. The changes in TEER between days 4 and 6 are shown as ΔTEER% (N = 4–8 per group). ∗ P < .05, ∗∗ P < .01, and ∗∗∗ P < .001. P values were determined by the ( A and D ) Kruskal–Wallis test followed by the Dunn multiple comparison test, and the ( B ) Mann–Whitney test.

    Article Snippet: Rabbit anti-human ALK1 antibody, rabbit anti-human CA1 antibody, rabbit anti-human OLFM4 antibody, and mouse/rabbit IgG VisUCyte horseradish-peroxidase polymer antibody were purchased from Sigma-Aldrich (St. Louis, MO) (HPA007041), Novus Biologicals (Littleton, CO) (NBP1-88191), Cell Signaling Technology (Beverly, MA) (14369S), and R&D Systems (VC002-025).

    Techniques: Expressing, Derivative Assay, Cell Culture, Isolation, Permeability, MANN-WHITNEY

    Decreased colonic ALK1 is associated with a poor clinical outcome in CD patients. ( A ) ALK1 expression was quantified in colonic biopsy samples obtained from NIBD controls (N = 10) and CD patients (N = 28) by qPCR. ( B ) ALK1 expression in the colonic mucosa of CD patients at the time of surgery and after surgery (N = 5 per group). ( C ) Percentages of CD patients who were diagnosed as CD before and after age 40 years in low-ALK1 (N = 15) and hi-ALK1 (N = 13) CD subsets. ( D ) Kaplan–Meier survival analysis to evaluate the impact of colonic ALK1 expression on endoscopic relapse in patients with CD (N = 12 for low-ALK1 and 9 for hi-ALK1 CD subgroups). ALK1 expression was normalized to GAPDH . ∗∗∗ P < .001. P values were determined by the ( A ) Mann–Whitney test, ( B ) Wilcoxon test, and ( D ) log-rank test.

    Journal: Cellular and Molecular Gastroenterology and Hepatology

    Article Title: Decreased Colonic Activin Receptor-Like Kinase 1 Disrupts Epithelial Barrier Integrity in Patients With Crohn’s Disease

    doi: 10.1016/j.jcmgh.2020.06.005

    Figure Lengend Snippet: Decreased colonic ALK1 is associated with a poor clinical outcome in CD patients. ( A ) ALK1 expression was quantified in colonic biopsy samples obtained from NIBD controls (N = 10) and CD patients (N = 28) by qPCR. ( B ) ALK1 expression in the colonic mucosa of CD patients at the time of surgery and after surgery (N = 5 per group). ( C ) Percentages of CD patients who were diagnosed as CD before and after age 40 years in low-ALK1 (N = 15) and hi-ALK1 (N = 13) CD subsets. ( D ) Kaplan–Meier survival analysis to evaluate the impact of colonic ALK1 expression on endoscopic relapse in patients with CD (N = 12 for low-ALK1 and 9 for hi-ALK1 CD subgroups). ALK1 expression was normalized to GAPDH . ∗∗∗ P < .001. P values were determined by the ( A ) Mann–Whitney test, ( B ) Wilcoxon test, and ( D ) log-rank test.

    Article Snippet: Rabbit anti-human ALK1 antibody, rabbit anti-human CA1 antibody, rabbit anti-human OLFM4 antibody, and mouse/rabbit IgG VisUCyte horseradish-peroxidase polymer antibody were purchased from Sigma-Aldrich (St. Louis, MO) (HPA007041), Novus Biologicals (Littleton, CO) (NBP1-88191), Cell Signaling Technology (Beverly, MA) (14369S), and R&D Systems (VC002-025).

    Techniques: Expressing, MANN-WHITNEY

    Clinical Characteristics at the Time of Sample Collection

    Journal: Cellular and Molecular Gastroenterology and Hepatology

    Article Title: Decreased Colonic Activin Receptor-Like Kinase 1 Disrupts Epithelial Barrier Integrity in Patients With Crohn’s Disease

    doi: 10.1016/j.jcmgh.2020.06.005

    Figure Lengend Snippet: Clinical Characteristics at the Time of Sample Collection

    Article Snippet: Rabbit anti-human ALK1 antibody, rabbit anti-human CA1 antibody, rabbit anti-human OLFM4 antibody, and mouse/rabbit IgG VisUCyte horseradish-peroxidase polymer antibody were purchased from Sigma-Aldrich (St. Louis, MO) (HPA007041), Novus Biologicals (Littleton, CO) (NBP1-88191), Cell Signaling Technology (Beverly, MA) (14369S), and R&D Systems (VC002-025).

    Techniques: Sampling, Activity Assay

    Correlations Between Explanatory Variables for the Risk of Surgery

    Journal: Cellular and Molecular Gastroenterology and Hepatology

    Article Title: Decreased Colonic Activin Receptor-Like Kinase 1 Disrupts Epithelial Barrier Integrity in Patients With Crohn’s Disease

    doi: 10.1016/j.jcmgh.2020.06.005

    Figure Lengend Snippet: Correlations Between Explanatory Variables for the Risk of Surgery

    Article Snippet: Rabbit anti-human ALK1 antibody, rabbit anti-human CA1 antibody, rabbit anti-human OLFM4 antibody, and mouse/rabbit IgG VisUCyte horseradish-peroxidase polymer antibody were purchased from Sigma-Aldrich (St. Louis, MO) (HPA007041), Novus Biologicals (Littleton, CO) (NBP1-88191), Cell Signaling Technology (Beverly, MA) (14369S), and R&D Systems (VC002-025).

    Techniques:

    Binomial Logistic Regression Analysis for Surgery

    Journal: Cellular and Molecular Gastroenterology and Hepatology

    Article Title: Decreased Colonic Activin Receptor-Like Kinase 1 Disrupts Epithelial Barrier Integrity in Patients With Crohn’s Disease

    doi: 10.1016/j.jcmgh.2020.06.005

    Figure Lengend Snippet: Binomial Logistic Regression Analysis for Surgery

    Article Snippet: Rabbit anti-human ALK1 antibody, rabbit anti-human CA1 antibody, rabbit anti-human OLFM4 antibody, and mouse/rabbit IgG VisUCyte horseradish-peroxidase polymer antibody were purchased from Sigma-Aldrich (St. Louis, MO) (HPA007041), Novus Biologicals (Littleton, CO) (NBP1-88191), Cell Signaling Technology (Beverly, MA) (14369S), and R&D Systems (VC002-025).

    Techniques:

    Primer Sequences for RT-qPCR

    Journal: Cellular and Molecular Gastroenterology and Hepatology

    Article Title: Decreased Colonic Activin Receptor-Like Kinase 1 Disrupts Epithelial Barrier Integrity in Patients With Crohn’s Disease

    doi: 10.1016/j.jcmgh.2020.06.005

    Figure Lengend Snippet: Primer Sequences for RT-qPCR

    Article Snippet: Rabbit anti-human ALK1 antibody, rabbit anti-human CA1 antibody, rabbit anti-human OLFM4 antibody, and mouse/rabbit IgG VisUCyte horseradish-peroxidase polymer antibody were purchased from Sigma-Aldrich (St. Louis, MO) (HPA007041), Novus Biologicals (Littleton, CO) (NBP1-88191), Cell Signaling Technology (Beverly, MA) (14369S), and R&D Systems (VC002-025).

    Techniques: