anti human cdx2  (Cell Signaling Technology Inc)

Journal: Phytotherapy Research

Article Title: Resveratrol inhibits bile acid‐induced gastric intestinal metaplasia via the PI3K / AKT / p‐FoxO4 signalling pathway

doi: 10.1002/ptr.6915

Figure Lengend Snippet: PCR primers sequences

Article Snippet: Primary antibodies against human CDX2 (#12306, 1:1000), Klf4 (#12173, 1:1000), Villin‐1 (#2369, 1:1000), cadherin‐17 (#42919, 1:1000), FoxO4 (#9472, 1:1000), AKT (#9272, 1:1000), phospho‐AKT Ser473 (#9271, 1:1000) and phospho‐AKT Thr308 (#9275, 1:1000) were purchased from Cell Signalling Technology (USA).

Techniques:

Journal: Phytotherapy Research

Article Title: Resveratrol inhibits bile acid‐induced gastric intestinal metaplasia via the PI3K / AKT / p‐FoxO4 signalling pathway

doi: 10.1002/ptr.6915

Figure Lengend Snippet: Potential FoxO4 binding site in the CDX2 promoter

Article Snippet: Primary antibodies against human CDX2 (#12306, 1:1000), Klf4 (#12173, 1:1000), Villin‐1 (#2369, 1:1000), cadherin‐17 (#42919, 1:1000), FoxO4 (#9472, 1:1000), AKT (#9272, 1:1000), phospho‐AKT Ser473 (#9271, 1:1000) and phospho‐AKT Thr308 (#9275, 1:1000) were purchased from Cell Signalling Technology (USA).

Techniques: Binding Assay, Sequencing

Journal: Phytotherapy Research

Article Title: Resveratrol inhibits bile acid‐induced gastric intestinal metaplasia via the PI3K / AKT / p‐FoxO4 signalling pathway

doi: 10.1002/ptr.6915

Figure Lengend Snippet: Resveratrol inhibits the expression of CDX2 and downstream intestinal markers in gastric cells and bile acid‐induced GIM. (A) The basic expression of CDX2 was measured at the protein (upper) and mRNA levels (lower) in GC cells and gastric epithelial cells (GES‐1). (B) The protein (upper) and mRNA (lower) expression levels of CDX2 were measured after CDCA stimulation (incubation time: 24 hr; dosage: 200 μM). The intestinal cancer cell line, HCT116, served as a positive control. (C) Cell immunofluorescence showed CDX2 expression in CDCA‐induced GIM (incubation time: 6 hr; dosage: 200 μM). Green: CDX2, Blue: Nuclei, red: Cytoskeletal protein. (D) Resveratrol down‐regulated CDX2 and Klf4 protein expression in a dose‐dependent manner in CDCA‐induced GIM (dosage of CDCA: 50 μM, Pre‐treatment for 24 hr) according to western blot analyses. (E–G) Resveratrol decreased the expression of CDX2 and downstream intestinal markers in a time‐ and dose‐dependent manner in AGS and MKN45 cells at the protein (upper) and mRNA (lower) levels. Unless otherwise specified, β‐actin, tubulin‐β and GAPDH were used as default western blot and RT‐PCR internal references. * p < .05; ** p < .01; *** p < .001; ns, not significant [Colour figure can be viewed at wileyonlinelibrary.com ]

Article Snippet: Primary antibodies against human CDX2 (#12306, 1:1000), Klf4 (#12173, 1:1000), Villin‐1 (#2369, 1:1000), cadherin‐17 (#42919, 1:1000), FoxO4 (#9472, 1:1000), AKT (#9272, 1:1000), phospho‐AKT Ser473 (#9271, 1:1000) and phospho‐AKT Thr308 (#9275, 1:1000) were purchased from Cell Signalling Technology (USA).

Techniques: Expressing, Incubation, Positive Control, Immunofluorescence, Western Blot, Reverse Transcription Polymerase Chain Reaction

Journal: Phytotherapy Research

Article Title: Resveratrol inhibits bile acid‐induced gastric intestinal metaplasia via the PI3K / AKT / p‐FoxO4 signalling pathway

doi: 10.1002/ptr.6915

Figure Lengend Snippet: FoxO4 was identified by the Cignal Finder 45‐Pathway Reporter Array and TranSignal Protein/DNA Array Kit and may combine directly with the promoter region of CDX2 as predicted by bioinformatics analysis. (A) After CDCA treatment of GES‐1 cells, changes in the activity of various transcription factors were determined using the TranSignal Protein/DNA Array Kit (fold regulation >2.0 and p value <.05). Red: increased fold regulation, blue: decreased fold regulation. Incubation time: 24 hr; dosage: 200 μM. For more detailed data, please see Tables and . (B) After resveratrol treatment of AGS cells, changes in the activity of various transcription factors were determined using the Cignal Finder 45‐Pathway Reporter Array (fold regulation >2.0 and p value <.05). Red: increased fold regulation, blue: decreased fold regulation. Incubation time: 24 hr; dosage: 200 μM. For more detailed data, please see Tables S2 and S4. (C) Transcription factors predicted by bioinformatics that could bind directly to the CDX2 promoter region. Green: CDX2, grey: transcription factors other than CDX2 with predicted direct binding to the CDX2 promoter. Red pentagram: FoxO4 or active FoxO4. (D) Relative activity change of the transcription factor FoxO4 after treatment with resveratrol and CDCA in gastric cells [Colour figure can be viewed at wileyonlinelibrary.com ]

Article Snippet: Primary antibodies against human CDX2 (#12306, 1:1000), Klf4 (#12173, 1:1000), Villin‐1 (#2369, 1:1000), cadherin‐17 (#42919, 1:1000), FoxO4 (#9472, 1:1000), AKT (#9272, 1:1000), phospho‐AKT Ser473 (#9271, 1:1000) and phospho‐AKT Thr308 (#9275, 1:1000) were purchased from Cell Signalling Technology (USA).

Techniques: DNA Array, Activity Assay, Incubation, Binding Assay

Journal: Phytotherapy Research

Article Title: Resveratrol inhibits bile acid‐induced gastric intestinal metaplasia via the PI3K / AKT / p‐FoxO4 signalling pathway

doi: 10.1002/ptr.6915

Figure Lengend Snippet: FoxO4 inhibits CDX2 expression by directly targeting the CDX2 promoter region. (A) Serially truncated CDX2 promoter constructs were cloned into pGL3‐luciferase reporter plasmids and transfected into GES‐1 cells. Four hours after transfection, the cells were treated with CDCA (200 μM) for 24 hr, and the relative luciferase activities were determined 72 hr later. (B) A ChIP assay demonstrated the direct binding of FoxO4 to the CDX2 promoter in GES‐1 cells. M: Marker. (C,D) qRT‐PCR of the ChIP products validated the binding capacity of FoxO4 to the CDX2 promoter. Means ± SEM of a representative experiment ( n = 3) performed in triplicate is shown. * p < .05; ** p < .01; *** p < .001; ns, not significant

Article Snippet: Primary antibodies against human CDX2 (#12306, 1:1000), Klf4 (#12173, 1:1000), Villin‐1 (#2369, 1:1000), cadherin‐17 (#42919, 1:1000), FoxO4 (#9472, 1:1000), AKT (#9272, 1:1000), phospho‐AKT Ser473 (#9271, 1:1000) and phospho‐AKT Thr308 (#9275, 1:1000) were purchased from Cell Signalling Technology (USA).

Techniques: Expressing, Construct, Clone Assay, Luciferase, Transfection, Binding Assay, Marker, Quantitative RT-PCR

Journal: Phytotherapy Research

Article Title: Resveratrol inhibits bile acid‐induced gastric intestinal metaplasia via the PI3K / AKT / p‐FoxO4 signalling pathway

doi: 10.1002/ptr.6915

Figure Lengend Snippet: CDX2 expression was negatively regulated by p‐FoxO4. (A‐B) GES‐1 and BGC823 cells were transfected with FoxO4 small interfering RNA (siFoxO4). With the decrease in p‐FoxO4 Ser262, the expression of CDX2 and intestinal markers increased at the protein (upper) and mRNA (lower) levels. (C) AGS cells were transfected with FoxO4 lentivirus. The absolute increase in p‐FoxO4 Ser262 could reduce the expression of CDX2 at the protein (upper) and mRNA (lower) levels

Article Snippet: Primary antibodies against human CDX2 (#12306, 1:1000), Klf4 (#12173, 1:1000), Villin‐1 (#2369, 1:1000), cadherin‐17 (#42919, 1:1000), FoxO4 (#9472, 1:1000), AKT (#9272, 1:1000), phospho‐AKT Ser473 (#9271, 1:1000) and phospho‐AKT Thr308 (#9275, 1:1000) were purchased from Cell Signalling Technology (USA).

Techniques: Expressing, Transfection, Small Interfering RNA

Journal: Phytotherapy Research

Article Title: Resveratrol inhibits bile acid‐induced gastric intestinal metaplasia via the PI3K / AKT / p‐FoxO4 signalling pathway

doi: 10.1002/ptr.6915

Figure Lengend Snippet: Resveratrol activated FoxO4 through the PI3K/AKT pathway. (A,B) The PI3K/AKT pathway inhibitor LY294004 partially blocked the effect of resveratrol on CDX2 at the protein level. (C) Relative protein levels of p‐AKT Ser473, p‐AKT Thr308, p‐FoxO4 Ser262 and CDX2 after treatment with the inhibitor LY294004. (D) AGS cells were transfected with siFoxO4. After a relative decrease in p‐FoxO4 resulted from the absolute reduction in FoxO4, the ability of resveratrol to reduce CDX2 weakened. (E) Relative protein levels of p‐FoxO4 Ser262 and CDX2 after siFoxO4 transfection. (F) GES‐1 cells were transfected with FoxO4 lentivirus. The relative increase in p‐FoxO4 suppressed the up‐regulation of CDX2 induced by CDCA. (G) Relative protein levels of p‐FoxO4 Ser262 and S197 and CDX2 after Lv‐FoxO4 transfection

Article Snippet: Primary antibodies against human CDX2 (#12306, 1:1000), Klf4 (#12173, 1:1000), Villin‐1 (#2369, 1:1000), cadherin‐17 (#42919, 1:1000), FoxO4 (#9472, 1:1000), AKT (#9272, 1:1000), phospho‐AKT Ser473 (#9271, 1:1000) and phospho‐AKT Thr308 (#9275, 1:1000) were purchased from Cell Signalling Technology (USA).

Techniques: Transfection

Journal: Phytotherapy Research

Article Title: Resveratrol inhibits bile acid‐induced gastric intestinal metaplasia via the PI3K / AKT / p‐FoxO4 signalling pathway

doi: 10.1002/ptr.6915

Figure Lengend Snippet: The PI3K/AKT/p‐FoxO4/CDX2 pathway is characteristic in normal and GIM tissue arrays. (A) Representative images of immunohistochemistry (IHC) staining for p‐FoxO4 Ser197 and CDX2 in normal tissues (upper) and IM tissues (lower) in 12 normal and 49 IM unpaired tissues. Scale bars: 20 μm. (Picture zoom 400 X; Embedded graphics 200X) (B) Relative protein levels of p‐FoxO4 S197 and CDX2 in normal (6.583 ± 0.570, n = 12; 1.417 ± 0.398, n = 12; Wilcoxon matched pairs, p value = .0005) and IM (2.367 ± 0.301, n = 49; 6.959 ± 0.464, n = 49; paired t test, p value <.0001) tissues detected by IHC. (C) Negative correlation between p‐FoxO4 Ser197 and CDX2 levels in normal and IM tissues ( n = 61, r = −.5216, p value <.0001) [Colour figure can be viewed at wileyonlinelibrary.com ]

Article Snippet: Primary antibodies against human CDX2 (#12306, 1:1000), Klf4 (#12173, 1:1000), Villin‐1 (#2369, 1:1000), cadherin‐17 (#42919, 1:1000), FoxO4 (#9472, 1:1000), AKT (#9272, 1:1000), phospho‐AKT Ser473 (#9271, 1:1000) and phospho‐AKT Thr308 (#9275, 1:1000) were purchased from Cell Signalling Technology (USA).

Techniques: Immunohistochemistry

Journal: Reproductive Sciences

Article Title: In Vitro Maturation of Fully Grown Mouse Antral Follicles in the Presence of 1 nM 2-Hydroxyestradiol Improves Oocytes’ Developmental Competence

doi: 10.1007/s43032-020-00276-6

Figure Lengend Snippet: Number ± SEM (*) of blastomeres forming the whole blastocyst, the inner cell mass (OCT4) or the trophectoderm (CDX2)

Article Snippet: Embryos were processed for immunolabelling using a rabbit polyclonal anti-human OCT4 (cat. N. ab19857, Abcam; diluted 1:400 in WS) or a rabbit anti-human CDX2 (cat. N. 3977, Cell Signaling Technology; diluted 1:100 in WS) antibodies.

Techniques: