esophagus rna  (Thermo Fisher)


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    Thermo Fisher esophagus rna
    Esophagus Rna, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 85 stars, based on 1 article reviews
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    esophagus rna - by Bioz Stars, 2020-09
    85/100 stars

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    Expressing:

    Article Title: Real-time PCR analysis of genes encoding tumor antigens in esophageal tumors and a cancer vaccine
    Article Snippet: .. Therefore, to enable consistent comparison of tumor samples to normal tissue, the average relative expression values for normal tissue biopsies from 8 patients was determined and compared with the relative expression level in purified esophagus RNA purchased from Ambion (Supplementary Figure 1). .. Gene expression was similar in all normal esophagus tissue samples; we therefore used the average relative expression in normal tissue biopsies from 8 patients as a baseline value to compare with tumor samples.

    Purification:

    Article Title: Real-time PCR analysis of genes encoding tumor antigens in esophageal tumors and a cancer vaccine
    Article Snippet: .. Therefore, to enable consistent comparison of tumor samples to normal tissue, the average relative expression values for normal tissue biopsies from 8 patients was determined and compared with the relative expression level in purified esophagus RNA purchased from Ambion (Supplementary Figure 1). .. Gene expression was similar in all normal esophagus tissue samples; we therefore used the average relative expression in normal tissue biopsies from 8 patients as a baseline value to compare with tumor samples.

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    Thermo Fisher kyse150 gfp cells
    Expression of OLC1 throughout the cell cycle. (A) Using serum starvation, <t>KYSE150/GFP</t> cells were synchronized at the G 0 phase. Following release, cells were collected every 2 for 24 h. The protein expression of OLC1, cyclin D1, cyclin A and actin was analyzed using western blot analysis. (B) Using 0.4 µg/ml nocodazole, KYSE150 /GFP cells were synchronized at the mitotic phase. Following release, cells were collected every 2 for 24 h. The protein expression of OLC1, cyclin B1, cyclin D1, cyclin A, cyclin E and actin was analyzed using western blot analysis. (C) Using reverse transcription polymerase chain reaction, the mRNA expression of OLC1 and GAPDH were detected. OLC1, overexpressed in lung cancer 1.
    Kyse150 Gfp Cells, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher sirna
    Knockdown of PKM2 altered the expression of apoptosis-related proteins. Eca109 or EC9706 cells (3×10 5 cells/well) were plated in a 12-well plate. PKM2 was knocked down in Eca109 and EC9706 cells by <t>siRNA</t> (50 nM final concentration for both PKM2-specific and NC siRNA). At 48 hours after <t>transfection,</t> the whole cell extracts were collected for western blot. Total RNA was extracted from cultured cells (Eca109 and EC9706) using Trizol reagent according to the manufacturer’s instructions. β-Actin and Bim mRNA were quantified in duplicate by SYBR Green 2-step, real-time reverse transcription polymerase chain reaction. (A) Caspase 9, caspase 3, cleaved caspase 3, Bcl-2, and Bim were detected by western blot. The results showed that knockout PKM2 activated caspase 3 and downregulated caspase 9. Bim protein expression was evidently increased. (B) Bim mRNA level was increased in depletion PKM2 esophageal cancer cells. PKM2, pyruvate kinase muscle isozyme 2; NC, normal control.
    Sirna, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 18683 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Thermo Fisher lrrc31
    <t>LRRC31</t> expression in esophageal epithelial cells a and CCL26 mRNA expression in primary esophageal epithelial cells treated with IL-13 (100 ng/mL) for 48 hours determined by qPCR analysis. b , Differentiated EPC2 esophageal epithelial cells were grown for 12 days in high-calcium media. Cells were brought to the ALI starting at day 7, and ±IL-13 treatment (100 ng/mL) started at day 8. c , Hematoxylin and eosin (H E)-stained sections of differentiated EPC2s at day 14. Stratum corneum (SC, pink layer) and stratum germinativum (SG, purple layer) are indicated. A representative experiment is shown (n = 3). Scale bar represents 20 μm. d , Normalized LRRC31 and CCL26 mRNA expression in differentiated EPC2s at day 14 determined by qPCR. A representative experiment is shown (n = 3). e , LRRC31 protein expression in differentiated EPC2s at day 14 determined by western blot analysis. Image shown is a representative experiment (n = 3); HSP90 (90 kD) loading control is shown. Left lane is the molecular weight ladder. Expression level of LRRC31 protein was quantified and normalized to the level of HSP90 protein. For a and d - h , data are represented as the mean ± SEM; * P
    Lrrc31, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 88/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher dsg1
    Impaired barrier function (IBF) in EoE can be replicated in <t>DSG1</t> deficient esophageal epithelial cells Representative electron micrographs of esophageal biopsies from healthy (NL) controls (n = 3) and patients with active EoE (n = 3). Arrowheads indicate the presence of dilated intercellular spaces (DIS) in EoE (A). TER (R T ) measurements from esophageal biopsies from healthy (NL) control and patients with active EoE (n = 6 and 9, respectively) (B). FITC-dextran flux assays from NL and active EoE esophageal biopsies (n = 6 and 4, respectively) (C). TER (R T ) measurements from NSC and DSG1 shRNA-transduced EPC2 cells following ALI differentiation (D). Kinetic analysis of FITC-dextran flux was also performed (E). Total FITC-dextran flux following 180 minutes are depicted in (F). Data in (D–F) are from two independent experiments performed in quadruplicate. All data are represented as the mean + SEM: *, p
    Dsg1, supplied by Thermo Fisher, 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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    Expression of OLC1 throughout the cell cycle. (A) Using serum starvation, KYSE150/GFP cells were synchronized at the G 0 phase. Following release, cells were collected every 2 for 24 h. The protein expression of OLC1, cyclin D1, cyclin A and actin was analyzed using western blot analysis. (B) Using 0.4 µg/ml nocodazole, KYSE150 /GFP cells were synchronized at the mitotic phase. Following release, cells were collected every 2 for 24 h. The protein expression of OLC1, cyclin B1, cyclin D1, cyclin A, cyclin E and actin was analyzed using western blot analysis. (C) Using reverse transcription polymerase chain reaction, the mRNA expression of OLC1 and GAPDH were detected. OLC1, overexpressed in lung cancer 1.

    Journal: Oncology Letters

    Article Title: Regulation of OLC1 protein expression by the anaphase-promoting complex

    doi: 10.3892/ol.2019.9881

    Figure Lengend Snippet: Expression of OLC1 throughout the cell cycle. (A) Using serum starvation, KYSE150/GFP cells were synchronized at the G 0 phase. Following release, cells were collected every 2 for 24 h. The protein expression of OLC1, cyclin D1, cyclin A and actin was analyzed using western blot analysis. (B) Using 0.4 µg/ml nocodazole, KYSE150 /GFP cells were synchronized at the mitotic phase. Following release, cells were collected every 2 for 24 h. The protein expression of OLC1, cyclin B1, cyclin D1, cyclin A, cyclin E and actin was analyzed using western blot analysis. (C) Using reverse transcription polymerase chain reaction, the mRNA expression of OLC1 and GAPDH were detected. OLC1, overexpressed in lung cancer 1.

    Article Snippet: Reverse transcription polymerase chain reaction (RT-PCR) Total RNA was extracted from treated KYSE150/GFP cells with TRIzol reagent (Thermo Fisher Scientific, Inc.), and RNA (6 µg) was reverse-transcribed according to the manufacturer's protocol (Super Script™ First-Strand Synthesis System for RT-PCR kit; Thermo Fisher Scientific, Inc., Waltham, MA, USA).

    Techniques: Expressing, Western Blot, Reverse Transcription Polymerase Chain Reaction

    OLC1 protein is degraded by the APC/c. (A) Analysis of the OLC1 protein sequence indicated that it contains a destruction box at the site of amino acids 12–20, which may be recognized by APC/c. (B) To investigate whether OLC1 may bind directly to the components of the APC/c, KYSE150/GFP-OLC1 cells were collected, lysed and subjected to IP. The presence of actin, OLC1 and APC2 in the immunocomplex were verified. (C) Different concentrations (2, 4 and 8 µg) of Myc-Cdh1/Cdc20 expression or mock vectors were transiently transfected into H1299 cells for 48 h. Then OLC1, Myc-Cdh1 and Myc-Cdc20 protein expression were evaluated. Actin was included as a loading control. Different concentrations (20, 50 and 100 nM) of (D) Cdh1 or (E) Cdc20 or mock siRNA were transiently transfected into H1299 cells for 48 h. Then OLC1, Cdh1 and Cdc20 protein expressions were evaluated. Actin was included as a loading control. OLC1, overexpressed in lung cancer 1; APC/c, anaphase-promoting cyclosome complex; IP, immunoprecipitation; Cdh1, cadherin 1; Cdc20, cell-division cycle protein 20; siRNA, small interfering RNA.

    Journal: Oncology Letters

    Article Title: Regulation of OLC1 protein expression by the anaphase-promoting complex

    doi: 10.3892/ol.2019.9881

    Figure Lengend Snippet: OLC1 protein is degraded by the APC/c. (A) Analysis of the OLC1 protein sequence indicated that it contains a destruction box at the site of amino acids 12–20, which may be recognized by APC/c. (B) To investigate whether OLC1 may bind directly to the components of the APC/c, KYSE150/GFP-OLC1 cells were collected, lysed and subjected to IP. The presence of actin, OLC1 and APC2 in the immunocomplex were verified. (C) Different concentrations (2, 4 and 8 µg) of Myc-Cdh1/Cdc20 expression or mock vectors were transiently transfected into H1299 cells for 48 h. Then OLC1, Myc-Cdh1 and Myc-Cdc20 protein expression were evaluated. Actin was included as a loading control. Different concentrations (20, 50 and 100 nM) of (D) Cdh1 or (E) Cdc20 or mock siRNA were transiently transfected into H1299 cells for 48 h. Then OLC1, Cdh1 and Cdc20 protein expressions were evaluated. Actin was included as a loading control. OLC1, overexpressed in lung cancer 1; APC/c, anaphase-promoting cyclosome complex; IP, immunoprecipitation; Cdh1, cadherin 1; Cdc20, cell-division cycle protein 20; siRNA, small interfering RNA.

    Article Snippet: Reverse transcription polymerase chain reaction (RT-PCR) Total RNA was extracted from treated KYSE150/GFP cells with TRIzol reagent (Thermo Fisher Scientific, Inc.), and RNA (6 µg) was reverse-transcribed according to the manufacturer's protocol (Super Script™ First-Strand Synthesis System for RT-PCR kit; Thermo Fisher Scientific, Inc., Waltham, MA, USA).

    Techniques: Sequencing, Expressing, Transfection, Immunoprecipitation, Small Interfering RNA

    OLC1 protein expression with CHX and MG132 treatment. (A) Following treatment with 100 µg/ml CHX, KYSE150/GFP cells were collected for western blot analysis of OLC1 protein expression at the indicated times. KYSE150/GFP and KYSE150/GFP-OLC1 cells were co-treated with CHX (100 µg/ml) and MG132 at (B) different concentrations (5, 10 and 20 µM MG132) or (C) for different lengths of time (4, 8, 12 and 16 h) with 20 µM MG132, with the DMSO, negative control and MG132-negative groups collected for analysis after 16 h of treatment. Results are representative of three independent experiments. (D) With (+) or without (−) treatment with 20 µM MG132, KYSE150/GFP-OLC1 cells were incubated for 16 h, collected and lysed for IP with an anti-OLC1 antibody. Ubiquitins were detected in the immunocomplex, and the presence of OLC1 was verified. IP with an anti-actin antibody was used as a negative control. OLC1, overexpressed in lung cancer 1; CHX, cycloheximide; IP, immunoprecipitation.

    Journal: Oncology Letters

    Article Title: Regulation of OLC1 protein expression by the anaphase-promoting complex

    doi: 10.3892/ol.2019.9881

    Figure Lengend Snippet: OLC1 protein expression with CHX and MG132 treatment. (A) Following treatment with 100 µg/ml CHX, KYSE150/GFP cells were collected for western blot analysis of OLC1 protein expression at the indicated times. KYSE150/GFP and KYSE150/GFP-OLC1 cells were co-treated with CHX (100 µg/ml) and MG132 at (B) different concentrations (5, 10 and 20 µM MG132) or (C) for different lengths of time (4, 8, 12 and 16 h) with 20 µM MG132, with the DMSO, negative control and MG132-negative groups collected for analysis after 16 h of treatment. Results are representative of three independent experiments. (D) With (+) or without (−) treatment with 20 µM MG132, KYSE150/GFP-OLC1 cells were incubated for 16 h, collected and lysed for IP with an anti-OLC1 antibody. Ubiquitins were detected in the immunocomplex, and the presence of OLC1 was verified. IP with an anti-actin antibody was used as a negative control. OLC1, overexpressed in lung cancer 1; CHX, cycloheximide; IP, immunoprecipitation.

    Article Snippet: Reverse transcription polymerase chain reaction (RT-PCR) Total RNA was extracted from treated KYSE150/GFP cells with TRIzol reagent (Thermo Fisher Scientific, Inc.), and RNA (6 µg) was reverse-transcribed according to the manufacturer's protocol (Super Script™ First-Strand Synthesis System for RT-PCR kit; Thermo Fisher Scientific, Inc., Waltham, MA, USA).

    Techniques: Expressing, Western Blot, Negative Control, Incubation, Immunoprecipitation

    Knockdown of PKM2 altered the expression of apoptosis-related proteins. Eca109 or EC9706 cells (3×10 5 cells/well) were plated in a 12-well plate. PKM2 was knocked down in Eca109 and EC9706 cells by siRNA (50 nM final concentration for both PKM2-specific and NC siRNA). At 48 hours after transfection, the whole cell extracts were collected for western blot. Total RNA was extracted from cultured cells (Eca109 and EC9706) using Trizol reagent according to the manufacturer’s instructions. β-Actin and Bim mRNA were quantified in duplicate by SYBR Green 2-step, real-time reverse transcription polymerase chain reaction. (A) Caspase 9, caspase 3, cleaved caspase 3, Bcl-2, and Bim were detected by western blot. The results showed that knockout PKM2 activated caspase 3 and downregulated caspase 9. Bim protein expression was evidently increased. (B) Bim mRNA level was increased in depletion PKM2 esophageal cancer cells. PKM2, pyruvate kinase muscle isozyme 2; NC, normal control.

    Journal: Cancer Research and Treatment : Official Journal of Korean Cancer Association

    Article Title: Effects and Mechanisms of Metformin on the Proliferation of Esophageal Cancer Cells In Vitro and In Vivo

    doi: 10.4143/crt.2015.485

    Figure Lengend Snippet: Knockdown of PKM2 altered the expression of apoptosis-related proteins. Eca109 or EC9706 cells (3×10 5 cells/well) were plated in a 12-well plate. PKM2 was knocked down in Eca109 and EC9706 cells by siRNA (50 nM final concentration for both PKM2-specific and NC siRNA). At 48 hours after transfection, the whole cell extracts were collected for western blot. Total RNA was extracted from cultured cells (Eca109 and EC9706) using Trizol reagent according to the manufacturer’s instructions. β-Actin and Bim mRNA were quantified in duplicate by SYBR Green 2-step, real-time reverse transcription polymerase chain reaction. (A) Caspase 9, caspase 3, cleaved caspase 3, Bcl-2, and Bim were detected by western blot. The results showed that knockout PKM2 activated caspase 3 and downregulated caspase 9. Bim protein expression was evidently increased. (B) Bim mRNA level was increased in depletion PKM2 esophageal cancer cells. PKM2, pyruvate kinase muscle isozyme 2; NC, normal control.

    Article Snippet: Eca109 or EC9706 cells were seeded in a 12-well plate (3×105 cells/well) and transfected with 50 nM siRNA using Lipofectamine 2000 Transfection Reagent (Thermo Fisher Scientific, Waltham, MA).

    Techniques: Expressing, Concentration Assay, Transfection, Western Blot, Cell Culture, SYBR Green Assay, Reverse Transcription Polymerase Chain Reaction, Knock-Out

    LRRC31 expression in esophageal epithelial cells a and CCL26 mRNA expression in primary esophageal epithelial cells treated with IL-13 (100 ng/mL) for 48 hours determined by qPCR analysis. b , Differentiated EPC2 esophageal epithelial cells were grown for 12 days in high-calcium media. Cells were brought to the ALI starting at day 7, and ±IL-13 treatment (100 ng/mL) started at day 8. c , Hematoxylin and eosin (H E)-stained sections of differentiated EPC2s at day 14. Stratum corneum (SC, pink layer) and stratum germinativum (SG, purple layer) are indicated. A representative experiment is shown (n = 3). Scale bar represents 20 μm. d , Normalized LRRC31 and CCL26 mRNA expression in differentiated EPC2s at day 14 determined by qPCR. A representative experiment is shown (n = 3). e , LRRC31 protein expression in differentiated EPC2s at day 14 determined by western blot analysis. Image shown is a representative experiment (n = 3); HSP90 (90 kD) loading control is shown. Left lane is the molecular weight ladder. Expression level of LRRC31 protein was quantified and normalized to the level of HSP90 protein. For a and d - h , data are represented as the mean ± SEM; * P

    Journal: Mucosal immunology

    Article Title: LRRC31 is induced by IL-13 and regulates kallikrein expression and barrier function in the esophageal epithelium

    doi: 10.1038/mi.2015.98

    Figure Lengend Snippet: LRRC31 expression in esophageal epithelial cells a and CCL26 mRNA expression in primary esophageal epithelial cells treated with IL-13 (100 ng/mL) for 48 hours determined by qPCR analysis. b , Differentiated EPC2 esophageal epithelial cells were grown for 12 days in high-calcium media. Cells were brought to the ALI starting at day 7, and ±IL-13 treatment (100 ng/mL) started at day 8. c , Hematoxylin and eosin (H E)-stained sections of differentiated EPC2s at day 14. Stratum corneum (SC, pink layer) and stratum germinativum (SG, purple layer) are indicated. A representative experiment is shown (n = 3). Scale bar represents 20 μm. d , Normalized LRRC31 and CCL26 mRNA expression in differentiated EPC2s at day 14 determined by qPCR. A representative experiment is shown (n = 3). e , LRRC31 protein expression in differentiated EPC2s at day 14 determined by western blot analysis. Image shown is a representative experiment (n = 3); HSP90 (90 kD) loading control is shown. Left lane is the molecular weight ladder. Expression level of LRRC31 protein was quantified and normalized to the level of HSP90 protein. For a and d - h , data are represented as the mean ± SEM; * P

    Article Snippet: EPC2s were transduced with shRNA targeting the coding sequence of LRRC31 or a NSC shRNA using the GIPZ lentiviral system (Thermo Fisher Scientific, Waltham, MA).

    Techniques: Expressing, Real-time Polymerase Chain Reaction, Staining, Western Blot, Molecular Weight

    KLK expression following LRRC31 gene-silencing in differentiated EPC2 cells EPC2 esophageal epithelial cells transduced with either non-silencing control (NSC) or LRRC31 short hairpin RNA (LRRC31 shRNA) lentiviral expression constructs were cultured as described in Figure 5b . a , Normalized LRRC31 and CCL26 mRNA expression in differentiated NSC and LRRC31 shRNA EPC2s at day 14 determined by qPCR analysis. Dashed line represents limit of detection. b , Fold change in LRRC31 mRNA expression following IL-13–treatment in differentiated NSC and LRRC31 shRNA EPC2s at day 14. c , H E-stained sections from differentiated NSC and LRRC31 shRNA EPC2s at day 14. d , Normalized KLK1, KLK5, KLK7, KLK11, and KLK13 mRNA expression in IL-13–treated differentiated NSC and LRRC31 shRNA EPC2 cells at day 14 determined by qPCR. e , TER measured across differentiated NSC and LRRC31 shRNA EPC2s at day 14. R T , resistance. For a - e , a representative experiment is shown (n = 3). For a , b , d , and e , data are represented as the mean ± SEM; NS, not significant; * P

    Journal: Mucosal immunology

    Article Title: LRRC31 is induced by IL-13 and regulates kallikrein expression and barrier function in the esophageal epithelium

    doi: 10.1038/mi.2015.98

    Figure Lengend Snippet: KLK expression following LRRC31 gene-silencing in differentiated EPC2 cells EPC2 esophageal epithelial cells transduced with either non-silencing control (NSC) or LRRC31 short hairpin RNA (LRRC31 shRNA) lentiviral expression constructs were cultured as described in Figure 5b . a , Normalized LRRC31 and CCL26 mRNA expression in differentiated NSC and LRRC31 shRNA EPC2s at day 14 determined by qPCR analysis. Dashed line represents limit of detection. b , Fold change in LRRC31 mRNA expression following IL-13–treatment in differentiated NSC and LRRC31 shRNA EPC2s at day 14. c , H E-stained sections from differentiated NSC and LRRC31 shRNA EPC2s at day 14. d , Normalized KLK1, KLK5, KLK7, KLK11, and KLK13 mRNA expression in IL-13–treated differentiated NSC and LRRC31 shRNA EPC2 cells at day 14 determined by qPCR. e , TER measured across differentiated NSC and LRRC31 shRNA EPC2s at day 14. R T , resistance. For a - e , a representative experiment is shown (n = 3). For a , b , d , and e , data are represented as the mean ± SEM; NS, not significant; * P

    Article Snippet: EPC2s were transduced with shRNA targeting the coding sequence of LRRC31 or a NSC shRNA using the GIPZ lentiviral system (Thermo Fisher Scientific, Waltham, MA).

    Techniques: Expressing, Transduction, shRNA, Construct, Cell Culture, Real-time Polymerase Chain Reaction, Staining

    Differentiated LRRC31-overexpressing EPC2 cell transcriptome a , Heat diagram representing RNA sequencing analysis of differentiated empty vector (control) and LRRC31-overexpressing EPC2s. Differentially expressed genes were identified by filtering on FPKM ≥ 1, moderated t-test with Benjamini-Hochberg False Discovery Rate (P

    Journal: Mucosal immunology

    Article Title: LRRC31 is induced by IL-13 and regulates kallikrein expression and barrier function in the esophageal epithelium

    doi: 10.1038/mi.2015.98

    Figure Lengend Snippet: Differentiated LRRC31-overexpressing EPC2 cell transcriptome a , Heat diagram representing RNA sequencing analysis of differentiated empty vector (control) and LRRC31-overexpressing EPC2s. Differentially expressed genes were identified by filtering on FPKM ≥ 1, moderated t-test with Benjamini-Hochberg False Discovery Rate (P

    Article Snippet: EPC2s were transduced with shRNA targeting the coding sequence of LRRC31 or a NSC shRNA using the GIPZ lentiviral system (Thermo Fisher Scientific, Waltham, MA).

    Techniques: RNA Sequencing Assay, Plasmid Preparation

    LRRC31 expression in the esophagus a , LRRC31 mRNA expression in esophageal biopsies of 16 subjects (6 normal controls [NL], 10 active eosinophilic esophagitis [EoE]) determined by RNA sequencing analysis. b , Normalized LRRC31 mRNA expression in esophageal biopsies (13 NL, 14 EoE) determined by quantitative PCR (qPCR) analysis. c , Normalized LRRC31 mRNA expression in esophageal biopsies (14 NL, 18 EoE) determined by microarray gene expression analysis. d , LRRC31 (61.5 kD) protein expression in esophageal biopsies (12 NL, 13 EoE) determined by western blot analysis. Image shown is a representative experiment; HSP90 (90 kD) loading control is shown. Expression level of LRRC31 protein was quantified and normalized to the level of HSP90 protein (mean normalized signal of all patients are graphed). For a - c , data points represent individual subjects. For a - d , data are represented as the mean ± SEM. * P

    Journal: Mucosal immunology

    Article Title: LRRC31 is induced by IL-13 and regulates kallikrein expression and barrier function in the esophageal epithelium

    doi: 10.1038/mi.2015.98

    Figure Lengend Snippet: LRRC31 expression in the esophagus a , LRRC31 mRNA expression in esophageal biopsies of 16 subjects (6 normal controls [NL], 10 active eosinophilic esophagitis [EoE]) determined by RNA sequencing analysis. b , Normalized LRRC31 mRNA expression in esophageal biopsies (13 NL, 14 EoE) determined by quantitative PCR (qPCR) analysis. c , Normalized LRRC31 mRNA expression in esophageal biopsies (14 NL, 18 EoE) determined by microarray gene expression analysis. d , LRRC31 (61.5 kD) protein expression in esophageal biopsies (12 NL, 13 EoE) determined by western blot analysis. Image shown is a representative experiment; HSP90 (90 kD) loading control is shown. Expression level of LRRC31 protein was quantified and normalized to the level of HSP90 protein (mean normalized signal of all patients are graphed). For a - c , data points represent individual subjects. For a - d , data are represented as the mean ± SEM. * P

    Article Snippet: EPC2s were transduced with shRNA targeting the coding sequence of LRRC31 or a NSC shRNA using the GIPZ lentiviral system (Thermo Fisher Scientific, Waltham, MA).

    Techniques: Expressing, RNA Sequencing Assay, Real-time Polymerase Chain Reaction, Microarray, Western Blot

    Expression of LRRC31 in human and murine tissue a , Normalized LRRC31 mRNA expression in various normal human tissues determined by microarray expression analysis. Data from GeneAtlas microarray dataset. 14 , 15 b , Normalized Lrrc31 mRNA expression in various normal murine tissues (C57BL/6) determined by qPCR analysis. c , Normalized LRRC31 mRNA expression in healthy human mucosal epithelium and epithelial cell lines determined by microarray expression analysis. Data from Barcode on Normal Tissue microarray dataset. 14 , 15 For a and c , data are represented as the mean ± SEM.

    Journal: Mucosal immunology

    Article Title: LRRC31 is induced by IL-13 and regulates kallikrein expression and barrier function in the esophageal epithelium

    doi: 10.1038/mi.2015.98

    Figure Lengend Snippet: Expression of LRRC31 in human and murine tissue a , Normalized LRRC31 mRNA expression in various normal human tissues determined by microarray expression analysis. Data from GeneAtlas microarray dataset. 14 , 15 b , Normalized Lrrc31 mRNA expression in various normal murine tissues (C57BL/6) determined by qPCR analysis. c , Normalized LRRC31 mRNA expression in healthy human mucosal epithelium and epithelial cell lines determined by microarray expression analysis. Data from Barcode on Normal Tissue microarray dataset. 14 , 15 For a and c , data are represented as the mean ± SEM.

    Article Snippet: EPC2s were transduced with shRNA targeting the coding sequence of LRRC31 or a NSC shRNA using the GIPZ lentiviral system (Thermo Fisher Scientific, Waltham, MA).

    Techniques: Expressing, Microarray, Real-time Polymerase Chain Reaction

    LRRC31 correlation with esophageal eosinophilia and disease-associated gene expression a , Normalized LRRC31 mRNA expression in esophageal biopsies of patients with untreated, fluticasone propionate (FP)–treated, or diet therapy–treated EoE (14 normal controls [NL], 18 active eosinophilic esophagitis [EoE], 24 EoE treatment responders [R], 12 EoE treatment non-responders [NR]) by microarray gene expression analysis. b , Correlation of biopsy eosinophil count per high-power field (hpf) and normalized LRRC31 mRNA expression in esophageal biopsies of patients with active EoE determined by microarray gene expression analysis. Dotted line represents 15 eosinophils/hpf, the diagnostic threshold for EoE. Correlation of normalized LRRC31 mRNA expression with normalized IL13 ( c ), CCL26 ( d ), and IL5 ( e ) mRNA expression in esophageal biopsies of patients with active EoE determined by qPCR analysis. For a , data points represent individual subjects and data are represented as the mean ± SEM. NS, not significant; * P

    Journal: Mucosal immunology

    Article Title: LRRC31 is induced by IL-13 and regulates kallikrein expression and barrier function in the esophageal epithelium

    doi: 10.1038/mi.2015.98

    Figure Lengend Snippet: LRRC31 correlation with esophageal eosinophilia and disease-associated gene expression a , Normalized LRRC31 mRNA expression in esophageal biopsies of patients with untreated, fluticasone propionate (FP)–treated, or diet therapy–treated EoE (14 normal controls [NL], 18 active eosinophilic esophagitis [EoE], 24 EoE treatment responders [R], 12 EoE treatment non-responders [NR]) by microarray gene expression analysis. b , Correlation of biopsy eosinophil count per high-power field (hpf) and normalized LRRC31 mRNA expression in esophageal biopsies of patients with active EoE determined by microarray gene expression analysis. Dotted line represents 15 eosinophils/hpf, the diagnostic threshold for EoE. Correlation of normalized LRRC31 mRNA expression with normalized IL13 ( c ), CCL26 ( d ), and IL5 ( e ) mRNA expression in esophageal biopsies of patients with active EoE determined by qPCR analysis. For a , data points represent individual subjects and data are represented as the mean ± SEM. NS, not significant; * P

    Article Snippet: EPC2s were transduced with shRNA targeting the coding sequence of LRRC31 or a NSC shRNA using the GIPZ lentiviral system (Thermo Fisher Scientific, Waltham, MA).

    Techniques: Expressing, Microarray, Diagnostic Assay, Real-time Polymerase Chain Reaction

    Epithelial barrier function in differentiated LRRC31-overexpressing EPC2 cells EPC2 esophageal epithelial cells transduced with either empty vector (control) or flag-epitope tagged LRRC31 lentiviral expression constructs were cultured as described in Figure 5b . a , Normalized LRRC31 , CCL26 , and KRT10 mRNA expression in differentiated control and LRRC31-overexpressing (LRRC31) EPC2s at day 14 determined by qPCR analysis. A representative experiment shown (n = 3). b , LRRC31 (61.5 kD) protein expression in differentiated control and LRRC31-overexpressing EPC2s at day 14 determined by western blot analysis. Image shown is a representative experiment (n = 3); HSP90 (90 kD) loading control is shown. Left lane is the molecular weight ladder. c , H E staining of differentiated control and LRRC31-overexpressing EPC2s at day 14. Scale bar represents 20 μm. A representative experiment is shown (n = 3). d , Cross-sectional area of H E stained sections from differentiated control and LRRC31-overexpressing EPC2s at day 14. A representative experiment is shown (n = 3). e , TER measured across differentiated control and LRRC31-overexpressing EPC2s at day 14. A representative experiment is shown (n = 3). R T , resistance. f , FITC-dextran (3-5 kD) paracellular flux measured at 3 hours after FITC-dextran was added to luminal surface of differentiated control and LRR31-overexpressing EPC2s at day 14. A representative experiment is shown (n = 3). For a and d - f , data are represented as the mean ± SEM; NS, not significant; * P

    Journal: Mucosal immunology

    Article Title: LRRC31 is induced by IL-13 and regulates kallikrein expression and barrier function in the esophageal epithelium

    doi: 10.1038/mi.2015.98

    Figure Lengend Snippet: Epithelial barrier function in differentiated LRRC31-overexpressing EPC2 cells EPC2 esophageal epithelial cells transduced with either empty vector (control) or flag-epitope tagged LRRC31 lentiviral expression constructs were cultured as described in Figure 5b . a , Normalized LRRC31 , CCL26 , and KRT10 mRNA expression in differentiated control and LRRC31-overexpressing (LRRC31) EPC2s at day 14 determined by qPCR analysis. A representative experiment shown (n = 3). b , LRRC31 (61.5 kD) protein expression in differentiated control and LRRC31-overexpressing EPC2s at day 14 determined by western blot analysis. Image shown is a representative experiment (n = 3); HSP90 (90 kD) loading control is shown. Left lane is the molecular weight ladder. c , H E staining of differentiated control and LRRC31-overexpressing EPC2s at day 14. Scale bar represents 20 μm. A representative experiment is shown (n = 3). d , Cross-sectional area of H E stained sections from differentiated control and LRRC31-overexpressing EPC2s at day 14. A representative experiment is shown (n = 3). e , TER measured across differentiated control and LRRC31-overexpressing EPC2s at day 14. A representative experiment is shown (n = 3). R T , resistance. f , FITC-dextran (3-5 kD) paracellular flux measured at 3 hours after FITC-dextran was added to luminal surface of differentiated control and LRR31-overexpressing EPC2s at day 14. A representative experiment is shown (n = 3). For a and d - f , data are represented as the mean ± SEM; NS, not significant; * P

    Article Snippet: EPC2s were transduced with shRNA targeting the coding sequence of LRRC31 or a NSC shRNA using the GIPZ lentiviral system (Thermo Fisher Scientific, Waltham, MA).

    Techniques: Transduction, Plasmid Preparation, FLAG-tag, Expressing, Construct, Cell Culture, Real-time Polymerase Chain Reaction, Western Blot, Molecular Weight, Staining

    Identification of LRRC31 Venn diagram depicting genes differentially expressed ≥ 1.5 fold in the esophagus in EoE (2789 genes) and in IL-13 treated esophageal epithelial cells (634 genes) by microarray gene expression analysis. Genes overlapping between these two data sets were identified (318 genes). LRRC31 is in bold and was increased 25 fold.

    Journal: Mucosal immunology

    Article Title: LRRC31 is induced by IL-13 and regulates kallikrein expression and barrier function in the esophageal epithelium

    doi: 10.1038/mi.2015.98

    Figure Lengend Snippet: Identification of LRRC31 Venn diagram depicting genes differentially expressed ≥ 1.5 fold in the esophagus in EoE (2789 genes) and in IL-13 treated esophageal epithelial cells (634 genes) by microarray gene expression analysis. Genes overlapping between these two data sets were identified (318 genes). LRRC31 is in bold and was increased 25 fold.

    Article Snippet: EPC2s were transduced with shRNA targeting the coding sequence of LRRC31 or a NSC shRNA using the GIPZ lentiviral system (Thermo Fisher Scientific, Waltham, MA).

    Techniques: Microarray, Expressing

    Function of LRRC31 in the esophageal epithelium IL-13 induces LRRC31 expression in esophageal epithelial cells that inhibits specific KLKs to increase barrier function.

    Journal: Mucosal immunology

    Article Title: LRRC31 is induced by IL-13 and regulates kallikrein expression and barrier function in the esophageal epithelium

    doi: 10.1038/mi.2015.98

    Figure Lengend Snippet: Function of LRRC31 in the esophageal epithelium IL-13 induces LRRC31 expression in esophageal epithelial cells that inhibits specific KLKs to increase barrier function.

    Article Snippet: EPC2s were transduced with shRNA targeting the coding sequence of LRRC31 or a NSC shRNA using the GIPZ lentiviral system (Thermo Fisher Scientific, Waltham, MA).

    Techniques: Expressing

    Impaired barrier function (IBF) in EoE can be replicated in DSG1 deficient esophageal epithelial cells Representative electron micrographs of esophageal biopsies from healthy (NL) controls (n = 3) and patients with active EoE (n = 3). Arrowheads indicate the presence of dilated intercellular spaces (DIS) in EoE (A). TER (R T ) measurements from esophageal biopsies from healthy (NL) control and patients with active EoE (n = 6 and 9, respectively) (B). FITC-dextran flux assays from NL and active EoE esophageal biopsies (n = 6 and 4, respectively) (C). TER (R T ) measurements from NSC and DSG1 shRNA-transduced EPC2 cells following ALI differentiation (D). Kinetic analysis of FITC-dextran flux was also performed (E). Total FITC-dextran flux following 180 minutes are depicted in (F). Data in (D–F) are from two independent experiments performed in quadruplicate. All data are represented as the mean + SEM: *, p

    Journal: Mucosal immunology

    Article Title: Desmoglein-1 regulates esophageal epithelial barrier function and immune responses in eosinophilic esophagitis

    doi: 10.1038/mi.2013.90

    Figure Lengend Snippet: Impaired barrier function (IBF) in EoE can be replicated in DSG1 deficient esophageal epithelial cells Representative electron micrographs of esophageal biopsies from healthy (NL) controls (n = 3) and patients with active EoE (n = 3). Arrowheads indicate the presence of dilated intercellular spaces (DIS) in EoE (A). TER (R T ) measurements from esophageal biopsies from healthy (NL) control and patients with active EoE (n = 6 and 9, respectively) (B). FITC-dextran flux assays from NL and active EoE esophageal biopsies (n = 6 and 4, respectively) (C). TER (R T ) measurements from NSC and DSG1 shRNA-transduced EPC2 cells following ALI differentiation (D). Kinetic analysis of FITC-dextran flux was also performed (E). Total FITC-dextran flux following 180 minutes are depicted in (F). Data in (D–F) are from two independent experiments performed in quadruplicate. All data are represented as the mean + SEM: *, p

    Article Snippet: DSG1 knockdown EPC2 cells were transduced with shRNA targeting the last exon of DSG1 or a NSC shRNA using the GIPZ lentiviral system (Thermo Fisher Scientific, Rockford, IL, USA).

    Techniques: shRNA

    DSG1 deficiency increases periostin ( POSTN ) expression qPCR analysis of POSTN expression in non-silencing control (NSC) and DSG1 shRNA-transduced EPC2 cells following ALI differentiation (A) and in patient biopsies (same cohort as in Fig. 1D ) (B). Spearman correlation between esophageal expression of POSTN and DSG1 (from Fig. 1D ) in patients with active EoE (C). Data in (A) are representative of three independent experiments performed in duplicate and represented as the mean + SEM: * p

    Journal: Mucosal immunology

    Article Title: Desmoglein-1 regulates esophageal epithelial barrier function and immune responses in eosinophilic esophagitis

    doi: 10.1038/mi.2013.90

    Figure Lengend Snippet: DSG1 deficiency increases periostin ( POSTN ) expression qPCR analysis of POSTN expression in non-silencing control (NSC) and DSG1 shRNA-transduced EPC2 cells following ALI differentiation (A) and in patient biopsies (same cohort as in Fig. 1D ) (B). Spearman correlation between esophageal expression of POSTN and DSG1 (from Fig. 1D ) in patients with active EoE (C). Data in (A) are representative of three independent experiments performed in duplicate and represented as the mean + SEM: * p

    Article Snippet: DSG1 knockdown EPC2 cells were transduced with shRNA targeting the last exon of DSG1 or a NSC shRNA using the GIPZ lentiviral system (Thermo Fisher Scientific, Rockford, IL, USA).

    Techniques: Expressing, Real-time Polymerase Chain Reaction, shRNA

    Loss of DSG1 promotes epithelial pro-inflammatory transcriptional responses Heatmap of 63 transcripts with differential expression ( p

    Journal: Mucosal immunology

    Article Title: Desmoglein-1 regulates esophageal epithelial barrier function and immune responses in eosinophilic esophagitis

    doi: 10.1038/mi.2013.90

    Figure Lengend Snippet: Loss of DSG1 promotes epithelial pro-inflammatory transcriptional responses Heatmap of 63 transcripts with differential expression ( p

    Article Snippet: DSG1 knockdown EPC2 cells were transduced with shRNA targeting the last exon of DSG1 or a NSC shRNA using the GIPZ lentiviral system (Thermo Fisher Scientific, Rockford, IL, USA).

    Techniques: Expressing

    Model of DSG1 dysregulation in EoE pathogenesis Downregulation of DSG1 by select Th2 cytokines (e.g. IL-13) results in impaired barrier function (IBF) and increased antigen exposure as well as the expression of pro-allergic mediators including periostin ( POSTN ) , forming a pathogenic cycle to further exacerbate allergic inflammation. Inset shows H E staining of inflamed esophageal mucosa with dilated intercellular spaces (DIS) (arrowheads) and eosinophilic infiltration (arrows).

    Journal: Mucosal immunology

    Article Title: Desmoglein-1 regulates esophageal epithelial barrier function and immune responses in eosinophilic esophagitis

    doi: 10.1038/mi.2013.90

    Figure Lengend Snippet: Model of DSG1 dysregulation in EoE pathogenesis Downregulation of DSG1 by select Th2 cytokines (e.g. IL-13) results in impaired barrier function (IBF) and increased antigen exposure as well as the expression of pro-allergic mediators including periostin ( POSTN ) , forming a pathogenic cycle to further exacerbate allergic inflammation. Inset shows H E staining of inflamed esophageal mucosa with dilated intercellular spaces (DIS) (arrowheads) and eosinophilic infiltration (arrows).

    Article Snippet: DSG1 knockdown EPC2 cells were transduced with shRNA targeting the last exon of DSG1 or a NSC shRNA using the GIPZ lentiviral system (Thermo Fisher Scientific, Rockford, IL, USA).

    Techniques: Expressing, Staining

    Loss of DSG1 protein expression in EoE Immunofluorescence (A) or immunohistochemical staining (B) of esophageal biopsy sections from controls (NL) and patients with active EoE. In (A), DSG1 (upper panel, in red) and DSG3 (lower panel, in green) are shown. Nuclei are stained with DAPI (blue). In (B), DSG1 (upper panel, in brown) and E-cadherin (lower panel, in brown) are shown. Dashed lines in (A) and (B) indicate the basal epithelial layer. Images in are representative of 4 patients per group.

    Journal: Mucosal immunology

    Article Title: Desmoglein-1 regulates esophageal epithelial barrier function and immune responses in eosinophilic esophagitis

    doi: 10.1038/mi.2013.90

    Figure Lengend Snippet: Loss of DSG1 protein expression in EoE Immunofluorescence (A) or immunohistochemical staining (B) of esophageal biopsy sections from controls (NL) and patients with active EoE. In (A), DSG1 (upper panel, in red) and DSG3 (lower panel, in green) are shown. Nuclei are stained with DAPI (blue). In (B), DSG1 (upper panel, in brown) and E-cadherin (lower panel, in brown) are shown. Dashed lines in (A) and (B) indicate the basal epithelial layer. Images in are representative of 4 patients per group.

    Article Snippet: DSG1 knockdown EPC2 cells were transduced with shRNA targeting the last exon of DSG1 or a NSC shRNA using the GIPZ lentiviral system (Thermo Fisher Scientific, Rockford, IL, USA).

    Techniques: Expressing, Immunofluorescence, Immunohistochemistry, Staining

    Differentiation of esophageal epithelial cells at the air-liquid interface (ALI) H E-stained sections of EPC2 cells grown in submerged cultures or differentiated at the ALI (A). qPCR analysis of desmoglein-1 ( DSG1 ) (B) and keratin 10 ( KRT10 ) (C) expression in submerged or ALI-differentiated EPC2 cells. Data are representative of 4 experiments performed in duplicate and are represented as the mean + SEM: ** p

    Journal: Mucosal immunology

    Article Title: Desmoglein-1 regulates esophageal epithelial barrier function and immune responses in eosinophilic esophagitis

    doi: 10.1038/mi.2013.90

    Figure Lengend Snippet: Differentiation of esophageal epithelial cells at the air-liquid interface (ALI) H E-stained sections of EPC2 cells grown in submerged cultures or differentiated at the ALI (A). qPCR analysis of desmoglein-1 ( DSG1 ) (B) and keratin 10 ( KRT10 ) (C) expression in submerged or ALI-differentiated EPC2 cells. Data are representative of 4 experiments performed in duplicate and are represented as the mean + SEM: ** p

    Article Snippet: DSG1 knockdown EPC2 cells were transduced with shRNA targeting the last exon of DSG1 or a NSC shRNA using the GIPZ lentiviral system (Thermo Fisher Scientific, Rockford, IL, USA).

    Techniques: Staining, Real-time Polymerase Chain Reaction, Expressing

    Loss of DSG1 reduces esophageal epithelial cell adhesion qPCR analysis of DSG1 (A) and DSG3 (B) in ALI-differentiated EPC2 cells stably transduced with non-silencing control (NSC) or DSG1 shRNA. H E-stained sections from stably transduced cells differentiated at the ALI (C). H E-stained sections from EPC2 cells exposed to the air interface and treated with 10 μg/mL ETA (WT) or the S195A inactive mutant for 24 h (D). Arrows (C–D) indicate cell separation within the suprabasal epithelium. Cytospins from NSC or DSG1 shRNA-transduced EPC2 cells following dispase adhesion assays (E) and quantification of dissociated cell clusters are shown (F). Images in (C–E) are representative of 4–5 experiments performed in duplicate. Data in (A–B) and (E) are from 3 experiments performed in duplicate and are represented as the mean + SEM: NS (not significant), * p

    Journal: Mucosal immunology

    Article Title: Desmoglein-1 regulates esophageal epithelial barrier function and immune responses in eosinophilic esophagitis

    doi: 10.1038/mi.2013.90

    Figure Lengend Snippet: Loss of DSG1 reduces esophageal epithelial cell adhesion qPCR analysis of DSG1 (A) and DSG3 (B) in ALI-differentiated EPC2 cells stably transduced with non-silencing control (NSC) or DSG1 shRNA. H E-stained sections from stably transduced cells differentiated at the ALI (C). H E-stained sections from EPC2 cells exposed to the air interface and treated with 10 μg/mL ETA (WT) or the S195A inactive mutant for 24 h (D). Arrows (C–D) indicate cell separation within the suprabasal epithelium. Cytospins from NSC or DSG1 shRNA-transduced EPC2 cells following dispase adhesion assays (E) and quantification of dissociated cell clusters are shown (F). Images in (C–E) are representative of 4–5 experiments performed in duplicate. Data in (A–B) and (E) are from 3 experiments performed in duplicate and are represented as the mean + SEM: NS (not significant), * p

    Article Snippet: DSG1 knockdown EPC2 cells were transduced with shRNA targeting the last exon of DSG1 or a NSC shRNA using the GIPZ lentiviral system (Thermo Fisher Scientific, Rockford, IL, USA).

    Techniques: Real-time Polymerase Chain Reaction, Stable Transfection, Transduction, shRNA, Staining, Mutagenesis

    IL-13 downregulates DSG1 and promotes IBF in esophageal epithelial cells H E-stained sections of EPC2 cells differentiated at the ALI in the presence of 0 (untreated), 10, or 100 ng/mL IL-13 (A). Arrows indicate a cell separation within to the suprabasal epithelium. Images are representative of 3 experiments performed in duplicate. Expression levels of DSG1 (B) and KRT10 (C) were measured by qPCR in submerged or ALI-differentiated EPC2 cells in the absence (0 ng/mL) or presence of IL-13 (10 or 100 ng/mL). TER (R T ) measurements on EPC2 cells at 0, 3, and 5 days following differentiation at the ALI in the absence (untreated) or presence of IL-13 (100 ng/mL) (D). Data are from 3 experiments performed in duplicate and are represented as the mean + SEM: NS (not significant), ** p

    Journal: Mucosal immunology

    Article Title: Desmoglein-1 regulates esophageal epithelial barrier function and immune responses in eosinophilic esophagitis

    doi: 10.1038/mi.2013.90

    Figure Lengend Snippet: IL-13 downregulates DSG1 and promotes IBF in esophageal epithelial cells H E-stained sections of EPC2 cells differentiated at the ALI in the presence of 0 (untreated), 10, or 100 ng/mL IL-13 (A). Arrows indicate a cell separation within to the suprabasal epithelium. Images are representative of 3 experiments performed in duplicate. Expression levels of DSG1 (B) and KRT10 (C) were measured by qPCR in submerged or ALI-differentiated EPC2 cells in the absence (0 ng/mL) or presence of IL-13 (10 or 100 ng/mL). TER (R T ) measurements on EPC2 cells at 0, 3, and 5 days following differentiation at the ALI in the absence (untreated) or presence of IL-13 (100 ng/mL) (D). Data are from 3 experiments performed in duplicate and are represented as the mean + SEM: NS (not significant), ** p

    Article Snippet: DSG1 knockdown EPC2 cells were transduced with shRNA targeting the last exon of DSG1 or a NSC shRNA using the GIPZ lentiviral system (Thermo Fisher Scientific, Rockford, IL, USA).

    Techniques: Staining, Expressing, Real-time Polymerase Chain Reaction

    Specific reduction in desmoglein-1 (DSG1) gene expression in EoE Heatmap depicting expression levels of desmogleins 1–4 ( DSG1–4 ) (A) and individual FPKM values for DSG1 and DSG3 (B-C) from RNA sequencing of esophageal biopsies from 10 patients with active EoE versus 6 healthy controls (NL). Quantitative PCR (qPCR) analysis of DSG1 expression in esophageal biopsies from NL (n = 25) and patients with active EoE (n = 39) (D). qPCR analysis of DSG1 expression in esophageal biopsies from NL (n = 11) and patients with inactive (n = 10) or active (n = 13) EoE (E). Data are represented as the median + interquartile range: NS (not significant), *** p

    Journal: Mucosal immunology

    Article Title: Desmoglein-1 regulates esophageal epithelial barrier function and immune responses in eosinophilic esophagitis

    doi: 10.1038/mi.2013.90

    Figure Lengend Snippet: Specific reduction in desmoglein-1 (DSG1) gene expression in EoE Heatmap depicting expression levels of desmogleins 1–4 ( DSG1–4 ) (A) and individual FPKM values for DSG1 and DSG3 (B-C) from RNA sequencing of esophageal biopsies from 10 patients with active EoE versus 6 healthy controls (NL). Quantitative PCR (qPCR) analysis of DSG1 expression in esophageal biopsies from NL (n = 25) and patients with active EoE (n = 39) (D). qPCR analysis of DSG1 expression in esophageal biopsies from NL (n = 11) and patients with inactive (n = 10) or active (n = 13) EoE (E). Data are represented as the median + interquartile range: NS (not significant), *** p

    Article Snippet: DSG1 knockdown EPC2 cells were transduced with shRNA targeting the last exon of DSG1 or a NSC shRNA using the GIPZ lentiviral system (Thermo Fisher Scientific, Rockford, IL, USA).

    Techniques: Expressing, RNA Sequencing Assay, Real-time Polymerase Chain Reaction