Journal: Oncotarget

Article Title: Positive feedback loop of hepatoma-derived growth factor and β-catenin promotes carcinogenesis of colorectal cancer

doi:

Figure Lengend Snippet: A. - B. β-catenin knockdown significantly suppressed HDGF mRNA expression in HCT116 A. and HT29 B. cells by real-time PCR analysis, respectively; C. - E. β-catenin knockdown inhibited HDGF protein expression in HCT116 cells C. and mainly inhibited nuclear HDGF, β-catenin, c-Myc, cyclin D1, MMP9 and phos-GSK-3β (Ser9) protein expression in HCT116 D. and HT29 E. cells; F. Recombinant Wnt3a and DKK1 increased and decreased HDGF and β-catenin expression in HCT116 cells, respectively; G. - I. Recombinant Wnt3a and DKK1 increased and decreased nuclear and cytoplasmic HDGF, β-catenin, c-Myc, cyclin D1, MMP9 and phos-GSK-3β (Ser9) protein expression in HCT116 G. , I. and LOVO H. cells by Western blot analysis, respectively.

Article Snippet: To further verify the effect of β-catenin on HDGF expression in CRC cells, HDGF and β-catenin protein expressions in HCT116 were induced by 100ng/ml human recombinant Wnt3a (R&D SYSTEMS) and inhibited by 200ng/ml human recombinant DKK1 (R&D SYSTEMS) for 48 hours by Western blot analysis, respectively (Figure ).

Techniques: Knockdown, Expressing, Real-time Polymerase Chain Reaction, Recombinant, Western Blot

Journal: BioMed Research International

Article Title: Designation of a Novel DKK1 Multiepitope DNA Vaccine and Inhibition of Bone Loss in Collagen-Induced Arthritic Mice

doi: 10.1155/2015/765490

Figure Lengend Snippet: Designation of the DNA vaccine.(a) B cell epitope scanning of human DKK1 was performed with the software DNASTAR 7.1. (b) The affinity of epitopes was measured by indirect ELISA. (c) The separated epitopes were immunized BALB/c mice and the immunogenicity of epitopes was measured by sandwich ELISA. (d) The separated epitopes were injected to BALB/c mice for seven days. TRAP staining was performed to identify the mature osteoclasts. Magnification: 200x; data are expressed as the mean ± SEM.

Article Snippet: After blocking with 5% horse serum, the sections were incubated with a goat anti-human DKK1 antibody (5 μ g/mL, R&D Systems) overnight at 4°C.

Techniques: Software, Indirect ELISA, Immunopeptidomics, Sandwich ELISA, Injection, Staining

Journal: BioMed Research International

Article Title: Designation of a Novel DKK1 Multiepitope DNA Vaccine and Inhibition of Bone Loss in Collagen-Induced Arthritic Mice

doi: 10.1155/2015/765490

Figure Lengend Snippet: Construction of the DNA vaccine. (a) The maps of recombinant DKK1 DNA vaccine. (b) The amino acid sequence of human DKK1. (c) The recombinant amino acid sequence of DKK1 DNA vaccine. Red line, 110–144aa; green line, 153–181aa; orange line, 182–216aa; blue line, 228–253aa; black line, signal peptide; box, PADRE.

Article Snippet: After blocking with 5% horse serum, the sections were incubated with a goat anti-human DKK1 antibody (5 μ g/mL, R&D Systems) overnight at 4°C.

Techniques: Recombinant, Sequencing

Journal: BioMed Research International

Article Title: Designation of a Novel DKK1 Multiepitope DNA Vaccine and Inhibition of Bone Loss in Collagen-Induced Arthritic Mice

doi: 10.1155/2015/765490

Figure Lengend Snippet: Expression and immunogenicity of the DNA vaccine. (a–c) Expression of the multiepitope protein in vitro and in vivo were determined in cell culture supernatants by ELISA, cell lysates by Western blotting, and the muscles of mice by immunohistochemical analysis. (d-e) The DNA vaccine induced a specific IgG antibody against human DKK1. The titer and the end-point titer of the specific antibody were tested by ELISA. Bars indicate 100 μ m. Data are expressed as the mean ± SEM, * P < 0.05, *** P < 0.001.

Article Snippet: After blocking with 5% horse serum, the sections were incubated with a goat anti-human DKK1 antibody (5 μ g/mL, R&D Systems) overnight at 4°C.

Techniques: Expressing, Immunopeptidomics, In Vitro, In Vivo, Cell Culture, Enzyme-linked Immunosorbent Assay, Western Blot, Muscles, Immunohistochemical staining

Journal: Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research

Article Title: Differential gene expression in cultured osteoblasts and bone marrow stromal cells from patients with Paget's disease of bone.

doi: 10.1359/jbmr.061108

Figure Lengend Snippet: FIG. 3. Upregulation of Dkk1 expression in pagetic osteoblasts and BMSCs. (A) The relative expression of Dkk1 was determined by real-time PCR, and values are expressed relative to the mean level of expression in the nonpagetic groups, which have been normal- ized to 1. The number of samples was the same as in Fig. 2. (B) The concentration of secreted Dkk1 protein in conditioned media samples as determined by ELISA. The con- ditioned media samples were collected dur- ing the last 72 h of incubation from the cell cultures described above. Data in A and B are presented as mean ± SE. ap < 0.05 and bp < 0.01 vs. control. (C) Correlation be- tween levels of Dkk1 mRNA in cells and se- creted Dkk1 protein in conditioned media. Levels of Dkk1 mRNA are expressed rela- tive to the expression of ribosomal RNA. The correlation was calculated using Deming (model II) linear regression (r 0.88, p < 0.0001).

Article Snippet: Microtiter plates (Greiner) were coated with 100 l of anti-Dkk1 polyclonal antibody (R&D Systems) at a concentration of 1 g/ml in PBS, pH 7.2.

Techniques: Expressing, Real-time Polymerase Chain Reaction, Concentration Assay, Enzyme-linked Immunosorbent Assay, Incubation, Control

Journal: Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research

Article Title: Differential gene expression in cultured osteoblasts and bone marrow stromal cells from patients with Paget's disease of bone.

doi: 10.1359/jbmr.061108

Figure Lengend Snippet: FIG. 7. Schema showing possible effects and interactions of the changes in osteoblast gene expression shown in this study. Solid lines denote production of a factor by that cell, and broken lines indicate a regulatory influence of a factor. The overproduction of IL-1 and IL-6 by the pagetic osteoblast will result in osteoclast proliferation, leading to further increases in IL-6 levels in the bone marrow microenvironment. Increased pro- duction of Dkk1 by the pagetic osteoblast will further increase IL-6 levels and reduce osteoblast proliferation. This combination of effects could account for the development of lytic lesions in early phase Paget’s disease. Over time, both the excess of Dkk1 and of IL-6 will result in increased differentiation of osteoblasts, thus promoting mineralization. This could contribute to the development of sclerosis in longer-standing pagetic lesions.

Article Snippet: Microtiter plates (Greiner) were coated with 100 l of anti-Dkk1 polyclonal antibody (R&D Systems) at a concentration of 1 g/ml in PBS, pH 7.2.

Techniques: Gene Expression

Journal: BMC Complementary Medicine and Therapies

Article Title: Ophiopogon japonicus-derived plant exosomes: a promising strategy for blocking the progression of diabetic vasculopathy to myocardial infarction

doi: 10.1186/s12906-026-05310-9

Figure Lengend Snippet: Detection of apoptosis, telomerase and protein. ( A and B ) The detection of flow cytometry. ( C ) The assay of ELISA for Telomerase. ( D and E ) The detection of western blot for TNF-α, IL-1β, LIMA1, and DKK1. ( F and G ) The detection of western blot for β-Catenin and VEGFA. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, while “ns” indicates no statistical significance

Article Snippet: Western blot was used to detect the protein change of core genes, namely, TNF-α (Proteintech, 60291-1-lg, 1:2000), IL-1β (Proteintech, 16806-1-AP, 1:2000), LIMA1 (HUABIO, ER62400, 1:2000), DKK1 (Proteintech, 21112-1-AP, 1:5000), β-Catenin (Proteintech, 66379-1-lg, 1:5000), VEGFA (ABclonal, A0280, 1:500).

Techniques: Flow Cytometry, Enzyme-linked Immunosorbent Assay, Western Blot

Journal: Journal of Translational Medicine

Article Title: DKK1 loss promotes endometrial fibrosis via autophagy and exosome-mediated macrophage-to-myofibroblast transition

doi: 10.1186/s12967-024-05402-5

Figure Lengend Snippet: DKK1 was is decreased in endometria of IUA patients. ( A ) Next-generation sequencing was used to detect transcriptome differences between normal proliferating endometrial tissues and endometrium of patient with uterine adhesions. Figure A is a heat map. Figure ( B ) is a volcanic map. ( C ) KEGG signaling pathway analysis of differentially expressed genes. ( D ) Masson staining was used to detect the degree of endometrial fibrosis in normal endometrial and endometrium of patients with uterine adhesion. The expressions of DKK1, wnt5a, β-catenin, COL1, ATG7, LC3-II, LAMP1 and P62 in normal endometrium and endometrium of patients with uterine adhesion were detected by immunohistochemistry. Immunofluorescence detection of α-SMA (green) and CD68 (red) in normal endometrial and endometrial tissues of adhesion patients. (E)Western blot assay was used to detect the expression of DKK1, wnt5a, β-catenin, COL1, ATG7, LC3-II, LAMP1 and P62 in normal endometrium and endometrium in patients with uterine adhesions. Error bars represent the standard error. The symbols * and ** indicate p < 0.05 and 0.01, respectively. Scale bar: 5 μm

Article Snippet: The following mouse strains were used: Pgr iresCre/+ mouse < B6.129 S(Cg)-Pgr tm1.1(cre)Shah /AndJ (Stock No: 017915> ) purchased from Cyagen Biosciences (Suzhou, China) and Dkk1 flox/flox (Dkk1 f/f ) mouse provided by Cyagen Biosciences (Suzhou, China) [ , ].

Techniques: Next-Generation Sequencing, Staining, Immunohistochemistry, Immunofluorescence, Western Blot, Expressing

Journal: Journal of Translational Medicine

Article Title: DKK1 loss promotes endometrial fibrosis via autophagy and exosome-mediated macrophage-to-myofibroblast transition

doi: 10.1186/s12967-024-05402-5

Figure Lengend Snippet: Loss of DKK1 in endometrium of mouse exhibited a fibrotic phenotype. ( A ) The homozygotes of DKK1 conditional knockout mice(DKK1 fl/fl, Cre+/− ) were identified by PCR. ( B ) Next-generation sequencing was used to detect transcriptome differences between endometrial tissues of DKK1 fl/fl, Cre+/− mice and endometrial tissues of DKK1 fl/fl, Cre−/− mice. Figure B is a heat map. Figure ( C ) is a volcanic map. ( D ) KEGG signaling pathway analysis of differentially expressed genes. (E) Masson staining was used to detect the degree of endometrial fibrosis between endometrial tissues of DKK1 fl/fl, Cre+/− mice and endometrial tissues of DKK1 fl/fl, Cre−/− mice. The expressions of DKK1, wnt5a, β-catenin, COL1, ATG7, LC3-II, LAMP1 and P62 were detected by immunohistochemistry between endometrial tissues of DKK1 fl/fl, Cre+/− mice and endometrial tissues of DKK1 fl/fl, Cre−/− mice. Immunofluorescence detection of α-SMA (green) and F4/80 (red) between endometrial tissues of DKK1 fl/fl, Cre+/− mice and endometrial tissues of DKK1 fl/fl, Cre−/− mice. Error bars represent the standard error. The symbols * and ** indicate p < 0.05 and 0.01, respectively. Scale bar: 5 μm

Article Snippet: The following mouse strains were used: Pgr iresCre/+ mouse < B6.129 S(Cg)-Pgr tm1.1(cre)Shah /AndJ (Stock No: 017915> ) purchased from Cyagen Biosciences (Suzhou, China) and Dkk1 flox/flox (Dkk1 f/f ) mouse provided by Cyagen Biosciences (Suzhou, China) [ , ].

Techniques: Knock-Out, Next-Generation Sequencing, Staining, Immunohistochemistry, Immunofluorescence

Journal: Journal of Translational Medicine

Article Title: DKK1 loss promotes endometrial fibrosis via autophagy and exosome-mediated macrophage-to-myofibroblast transition

doi: 10.1186/s12967-024-05402-5

Figure Lengend Snippet: Dkk1 promoted autophagy of endometrial stromal cells. ( A ) After silencing DKK1 in primary endometrial stromal cells (PESC) and endometrial stromal cells immortalized cell lines (T-HESCS), western blot analysis was performed to detect the expression of DKK1,LC3-II, ATG7 and P62 proteins. ( B )After knock out of DKK1 in primary endometrial stromal cells (PESC) and endometrial stromal cells immortalized cell lines (T-HESCS), western blot analysis was performed to detect the expression of DKK1,LC3-II, ATG7 and P62 proteins. ( C ) After ectopic expression of DKK1 in primary endometrial stromal cells (PESC) and endometrial stromal cells immortalized cell lines (T-HESCS), western blot analysis was performed to detect the expression of DKK1,LC3-II, ATG7 and P62 proteins. Error bars represent the standard error. The symbols * and ** indicate p < 0.05 and 0.01, respectively

Article Snippet: The following mouse strains were used: Pgr iresCre/+ mouse < B6.129 S(Cg)-Pgr tm1.1(cre)Shah /AndJ (Stock No: 017915> ) purchased from Cyagen Biosciences (Suzhou, China) and Dkk1 flox/flox (Dkk1 f/f ) mouse provided by Cyagen Biosciences (Suzhou, China) [ , ].

Techniques: Western Blot, Expressing, Knock-Out

Journal: Journal of Translational Medicine

Article Title: DKK1 loss promotes endometrial fibrosis via autophagy and exosome-mediated macrophage-to-myofibroblast transition

doi: 10.1186/s12967-024-05402-5

Figure Lengend Snippet: DKK1 regulates Wnt/β-catenin and PI3K/AKT/mTOR pathways. ( A ) After knock out of DKK1 in primary endometrial stromal cells, next-generation sequencing was used to detect transcriptome differences between KO group and WT group. Figure ( A ) is a heat map. Figure ( B ) is a volcanic map. ( C ) KEGG signaling pathway analysis of differentially expressed genes. ( D ) After silencing Wnt5a in primary endometrial stromal cells (PESC) and endometrial stromal cells immortalized cell lines (T-HESCS), western blot analysis was performed to detect the expression of Wnt5a, LC3-II, ATG7 and P62 proteins. ( E ) After silencing TCF4 in primary endometrial stromal cells (PESC) and endometrial stromal cells immortalized cell lines (T-HESCS), western blot analysis was performed to detect the expression of TCF4, LC3-II, ATG7 and P62 proteins. ( F ) After knock out of DKK1 in primary endometrial stromal cells (PESC) and endometrial stromal cells immortalized cell lines (T-HESCS), western blot analysis was performed to detect the expression of WNT5A, β-catenin, p-akt, p38 and ATF4 proteins. ( G ) After ectopic expression of DKK1 in primary endometrial stromal cells (PESC) and endometrial stromal cells immortalized cell lines (T-HESCS), western blot analysis was performed to detect the expression of WNT5A, β-catenin, p-akt, p38 and ATF4 proteins. Error bars represent the standard error. The symbols * and ** indicate p < 0.05 and 0.01, respectively

Article Snippet: The following mouse strains were used: Pgr iresCre/+ mouse < B6.129 S(Cg)-Pgr tm1.1(cre)Shah /AndJ (Stock No: 017915> ) purchased from Cyagen Biosciences (Suzhou, China) and Dkk1 flox/flox (Dkk1 f/f ) mouse provided by Cyagen Biosciences (Suzhou, China) [ , ].

Techniques: Knock-Out, Next-Generation Sequencing, Western Blot, Expressing

Journal: Journal of Translational Medicine

Article Title: DKK1 loss promotes endometrial fibrosis via autophagy and exosome-mediated macrophage-to-myofibroblast transition

doi: 10.1186/s12967-024-05402-5

Figure Lengend Snippet: DKK1 ko promotes the release of exosomes. ( A ) The expression of TSG101 protein in primary endometrial stromal cells (PESC) was detected by western blot after DKK1 gene was knocked out. ( B ) primary endometrial stromal cells (PESC) with DKK1 overexpression were treated with bafa1. After 24 h, TSG101 protein was detected by western blot. Wild-type primary endometrial stromal cells (PESC) ( C , E ) and PESC with dkk1 gene knockout ( D and F )were used to detect exosome morphology and diameter. ( G ) Quantitative analysis of primary endometrial stromal cells (PESC)-derived exosomes by ELISA. Data are the means ± SEM of 3 assays. ( H ) The expression of exosomal markers TSG101 and HSPA8 was detected by western blotting. Error bars represent the standard error. The symbols * and ** indicate p < 0.05 and 0.01, respectively

Article Snippet: The following mouse strains were used: Pgr iresCre/+ mouse < B6.129 S(Cg)-Pgr tm1.1(cre)Shah /AndJ (Stock No: 017915> ) purchased from Cyagen Biosciences (Suzhou, China) and Dkk1 flox/flox (Dkk1 f/f ) mouse provided by Cyagen Biosciences (Suzhou, China) [ , ].

Techniques: Expressing, Western Blot, Over Expression, Gene Knockout, Derivative Assay, Enzyme-linked Immunosorbent Assay

Journal: Journal of Translational Medicine

Article Title: DKK1 loss promotes endometrial fibrosis via autophagy and exosome-mediated macrophage-to-myofibroblast transition

doi: 10.1186/s12967-024-05402-5

Figure Lengend Snippet: DKK1 exosomes promoted the transformation of macrophages into myofibroblasts. ( A - B ) Exosomes were extracted from Wild-type primary endometrial stromal cells (PESC) and PESC with DKK1 gene knockout. 72 h after exosomes were added to human macrophages, EdU assay was used to detect macrophage proliferation, and transwell assay was used to detect macrophage invasion status. The protein expressions of col1 and α-SMA were detected by Western blot. ( C - D ) Exosomes were extracted from Wild-type primary endometrial stromal cells (PESC) and PESC with DKK1 gene knockout. Cytokine antibody microarray was used to detect the expression of cytokines in exosomes derived from wild-type and DKK1 KO endometrial stromal cells. Knockout of DKK1-derived exosomes in endometrial stromal cells significantly increased the expression of IL-8 and IL-17 A. ( E ) elisa was used to detect the expression of IL-8 in exosomes derived from wild-type endometrial primary stromal cells or endometrial primary stromal cells with DKK1 gene knockout. ( F - G ) IL-8 was added to human macrophages for 72 h, EdU assay was used to detect macrophage proliferation, and transwell assay was used to detect macrophage invasion status. The protein expressions of col1 and α-SMA were detected by Western blot. Error bars represent the standard error. The symbols * and ** indicate p < 0.05 and 0.01, respectively. Scale bar: 5 μm

Article Snippet: The following mouse strains were used: Pgr iresCre/+ mouse < B6.129 S(Cg)-Pgr tm1.1(cre)Shah /AndJ (Stock No: 017915> ) purchased from Cyagen Biosciences (Suzhou, China) and Dkk1 flox/flox (Dkk1 f/f ) mouse provided by Cyagen Biosciences (Suzhou, China) [ , ].

Techniques: Transformation Assay, Gene Knockout, EdU Assay, Transwell Assay, Western Blot, Microarray, Expressing, Derivative Assay, Knock-Out, Enzyme-linked Immunosorbent Assay

Journal: Journal of Translational Medicine

Article Title: DKK1 loss promotes endometrial fibrosis via autophagy and exosome-mediated macrophage-to-myofibroblast transition

doi: 10.1186/s12967-024-05402-5

Figure Lengend Snippet: Autophagy activator inhibited endometrial fibrosis in DKK1 cko mice. ( A ) DKK1 fl/fl, Cre+/− mice were randomly divided into two groups. Rapamycin treatment group was given intraperitoneal injection of rapamycin(3 mg/kg/day). The control group was injected with equal volume of DMSO. Two weeks after administration of rapamycin, the mice were sacrificed. ( A - E ) Masson staining was used to detect the degree of endometrial fibrosis in rapamycin treatment group and control group. The expressions of DKK1, wnt5a, β-catenin, COL1, ATG7, LC3-II, LAMP1 and P62 in rapamycin treatment group and control group were detected by immunohistochemistry. Immunofluorescence detection of α-SMA (green) and F4/80 (red) in rapamycin treatment group and control group. Error bars represent the standard error. The symbols * and ** indicate p < 0.05 and 0.01, respectively. Scale bar: 5 μm

Article Snippet: The following mouse strains were used: Pgr iresCre/+ mouse < B6.129 S(Cg)-Pgr tm1.1(cre)Shah /AndJ (Stock No: 017915> ) purchased from Cyagen Biosciences (Suzhou, China) and Dkk1 flox/flox (Dkk1 f/f ) mouse provided by Cyagen Biosciences (Suzhou, China) [ , ].

Techniques: Injection, Control, Staining, Immunohistochemistry, Immunofluorescence