Journal: Scientific Reports

Article Title: Folic acid inhibits the Wnt/β-catenin pathway by upregulating DKK3 to exert anti-tumor effects in cervical squamous cell carcinoma

doi: 10.1038/s41598-025-32762-9

Figure Lengend Snippet: Establishment of DKK3-manipulated SiHa cell models with altered Wnt/β-catenin signaling activity. ( a , f ) Brightfield and fluorescence microscopy images after transfection with different lentivirus in SiHa cells cultured in medium (scale bar: 100 μm). ( b , g ) DKK3 mRNA expression in different treated cells ( n = 3). ( c - e , h - j ) Western blotting and corresponding quantitative expression analysis of DKK3 and β-catenin ( n = 3). Original blots are presented in Supplementary Fig. . * P < 0.05, ** P < 0.01, and *** P < 0.001 represent significant difference.

Article Snippet: Next, anti-DKK3 primary antibody (1:100 dilution; cat # 66758-1-lg, Proteintech, Wuhan, China) was applied to the sections for overnight incubation at 4 °C after a 30-min blocking step.

Techniques: Activity Assay, Fluorescence, Microscopy, Transfection, Cell Culture, Expressing, Western Blot

Journal: Scientific Reports

Article Title: Folic acid inhibits the Wnt/β-catenin pathway by upregulating DKK3 to exert anti-tumor effects in cervical squamous cell carcinoma

doi: 10.1038/s41598-025-32762-9

Figure Lengend Snippet: Cell proliferation, migration and invasion are affected by DKK3 expression in vitro. ( a , i ) CCK-8 assay detected the ability of cell proliferation ( n = 3). ( b - c , j - k ) Colony formation assay and count of different groups of SiHa cells( n = 3). ( d - e , l - m ) Representative wound healing assay images and analysis of different groups of SiHa cells ( n = 3). ( f - h , n - p ) Representative cell migration and invasion images and analysis of SiHa cells from different groups using transwell assay ( n = 3). * P < 0.05, ** P < 0.01, and *** P < 0.001 represent significant difference.

Article Snippet: Next, anti-DKK3 primary antibody (1:100 dilution; cat # 66758-1-lg, Proteintech, Wuhan, China) was applied to the sections for overnight incubation at 4 °C after a 30-min blocking step.

Techniques: Migration, Expressing, In Vitro, CCK-8 Assay, Colony Assay, Wound Healing Assay, Transwell Assay

Journal: Scientific Reports

Article Title: Folic acid inhibits the Wnt/β-catenin pathway by upregulating DKK3 to exert anti-tumor effects in cervical squamous cell carcinoma

doi: 10.1038/s41598-025-32762-9

Figure Lengend Snippet: FA inhibits cell viability, migration and invasion by upregulating DKK3 and reducing β-catenin protein abundance. ( a ) schematic illustration of the DKK3 regulation after cells treated with FA. ( b ) Cell viability of SiHa cells treated with different concentrations of FA ( n = 3). ( c )Cell viability of SiHa cells treated with different time course of FA ( n = 3). ( d ) Flow cytometry analysis for cell apoptosis of SiHa cells with different treatments ( n = 3). ( e ) Representative images for flow cytometry of SiHa cells with different treatments. ( f - g ) Representative wound healing assay images and analysis of FA treatment in SiHa cells ( n = 3). ( h ) Representative images of cell migration and invasion in SiHa cells treated with FA, using transwell assay. ( i - j ) Corresponding quantitative analysis of migration and invasion ( n = 3). (k) DKK3 mRNA levels in SiHa cells treated with different concentrations of FA ( n = 3). ( l – n ) Protein expression analysis of DKK3 and β‑catenin in SiHa cells under different FA treatments: representative western blot images (l) and corresponding quantification of DKK3 (m) and β‑catenin (n) ( n = 3). ( o ) Rescue experiments: DKK3 mRNA levels in DKK3‑knockdown SiHa cells treated with different concentrations of FA ( n = 3). ( p – r ) Rescue experiments: Protein expression analysis in DKK3‑knockdown SiHa cells under FA treatment: representative western blot images ( p ) and corresponding quantification of DKK3 ( q ) and β‑catenin ( r ) ( n = 3). Original blots are presented in Supplementary Fig. .* P < 0.05, ** P < 0.01, and *** P < 0.001 represent significant difference.

Article Snippet: Next, anti-DKK3 primary antibody (1:100 dilution; cat # 66758-1-lg, Proteintech, Wuhan, China) was applied to the sections for overnight incubation at 4 °C after a 30-min blocking step.

Techniques: Migration, Quantitative Proteomics, Flow Cytometry, Wound Healing Assay, Transwell Assay, Expressing, Western Blot

Journal: Discover Oncology

Article Title: Potential role of DKK3 and WIF1 in prostate cancer: bioinformatics and clinical analysis

doi: 10.1007/s12672-025-03488-x

Figure Lengend Snippet: The expression level of core genes. A DKK3 expression levels of the GSE69223 dataset and GSE46602 dataset; B WIF1 expression levels of the GSE69223 dataset and GSE46602 dataset; Compared with BPH group, * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: The primary antibodies used in this study included an anti-DKK3 polyclonal antibody (Cat. No. 10365-1-AP, Sangon Biotech, China), an anti-WIF1 rabbit polyclonal antibody (Cat. No. 53483-1, SAB Biotech, USA), and an anti-GAPDH monoclonal antibody (Cat. No. 60004-1-Ig, Sangon Biotech, China).

Techniques: Expressing

Journal: Discover Oncology

Article Title: Potential role of DKK3 and WIF1 in prostate cancer: bioinformatics and clinical analysis

doi: 10.1007/s12672-025-03488-x

Figure Lengend Snippet: Immune correlation analysis of DKK3

Article Snippet: The primary antibodies used in this study included an anti-DKK3 polyclonal antibody (Cat. No. 10365-1-AP, Sangon Biotech, China), an anti-WIF1 rabbit polyclonal antibody (Cat. No. 53483-1, SAB Biotech, USA), and an anti-GAPDH monoclonal antibody (Cat. No. 60004-1-Ig, Sangon Biotech, China).

Techniques:

Journal: Discover Oncology

Article Title: Potential role of DKK3 and WIF1 in prostate cancer: bioinformatics and clinical analysis

doi: 10.1007/s12672-025-03488-x

Figure Lengend Snippet: GSEA enrichment analysis of DKK3. A GO-BP functional annotation analysis was conducted for DKK3. B GO-CC functional annotation analysis was conducted for DKK3. C GO-MF functional annotation analysis was conducted for DKK3. D KEGG pathway analysis was performed for DKK3

Article Snippet: The primary antibodies used in this study included an anti-DKK3 polyclonal antibody (Cat. No. 10365-1-AP, Sangon Biotech, China), an anti-WIF1 rabbit polyclonal antibody (Cat. No. 53483-1, SAB Biotech, USA), and an anti-GAPDH monoclonal antibody (Cat. No. 60004-1-Ig, Sangon Biotech, China).

Techniques: Functional Assay

Journal: Discover Oncology

Article Title: Potential role of DKK3 and WIF1 in prostate cancer: bioinformatics and clinical analysis

doi: 10.1007/s12672-025-03488-x

Figure Lengend Snippet: Correlation analysis of DKK3 and WIF1 expression with CTRP drug sensitivity and mRNA expression. A Correlation analysis of DKK3 expression with CTRP drug sensitivity and mRNA expression. B Correlation analysis of WIF1 expression with CTRP drug sensitivity and mRNA expression

Article Snippet: The primary antibodies used in this study included an anti-DKK3 polyclonal antibody (Cat. No. 10365-1-AP, Sangon Biotech, China), an anti-WIF1 rabbit polyclonal antibody (Cat. No. 53483-1, SAB Biotech, USA), and an anti-GAPDH monoclonal antibody (Cat. No. 60004-1-Ig, Sangon Biotech, China).

Techniques: Expressing

Journal: Discover Oncology

Article Title: Potential role of DKK3 and WIF1 in prostate cancer: bioinformatics and clinical analysis

doi: 10.1007/s12672-025-03488-x

Figure Lengend Snippet: Expression of DKK3 and WIF1 in BPH and PCa. A Expression of DKK3 in BPH. B Expression of DKK3 in PCa. C Expression of WIF1 in BPH. D Expression of WIF1 in PCa. E The protein expression of DKK3 and WIF 1 in BPH and PCa. Compared with BPH group, * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: The primary antibodies used in this study included an anti-DKK3 polyclonal antibody (Cat. No. 10365-1-AP, Sangon Biotech, China), an anti-WIF1 rabbit polyclonal antibody (Cat. No. 53483-1, SAB Biotech, USA), and an anti-GAPDH monoclonal antibody (Cat. No. 60004-1-Ig, Sangon Biotech, China).

Techniques: Expressing

Journal: BMC cancer

Article Title: Targeting DKK3 to remodel tumor immune microenvironment and enhance cancer immunotherapy.

doi: 10.1186/s12885-025-14075-2

Figure Lengend Snippet: Fig. 1 High DKK3 expression is associated with worse patient survival across different cancer types. (A) DKK3 expression in normal adjacent tissues and tumors of patients from TCGA-PAAD, HNSC, THYM and DLBC. (B) The overall survival analysis of patients with high or low DKK3 expression from TCGA (pan-cancer), STAD, BLCA, GMB, HNSC and MESO. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 as calculated by log-rank test or unpaired Student’s t-test

Article Snippet: They received intraperitoneal injections of 10 mg/kg of a mouse monoclonal DKK3 antibody (DKK3-4.22, BE0385, Bio X Cell), a mouse functional PD-1 antibody (BE0146, Bio X Cell), or a combination of both.

Techniques: Expressing

Journal: BMC cancer

Article Title: Targeting DKK3 to remodel tumor immune microenvironment and enhance cancer immunotherapy.

doi: 10.1186/s12885-025-14075-2

Figure Lengend Snippet: Fig. 2 DKK3 expression is associated with immunosuppressive TME across different cancer types. (A) The interrelation of DKK3 expression with the infiltration percentage of CD8+ T cells in tumor samples from TCGA-BRCA, COAD, ESCA, LUSC and PAAD. (B) The interrelation of DKK3 expression with the infiltration percentage of Th1 cells in tumor samples from TCGA-BRCA, COAD, KICH, LGG and STAD. (C) The interrelation of DKK3 expression with the infiltration percentage of Treg cells in tumor samples from TCGA-BLCA, COAD, LUAD, PAAD and PRAD. (D) The interrelation of DKK3 expression with the infiltration percentage of M2 macrophages in tumor samples from TCGA-BLCA, COAD, KICH, READ and TGCT. (E) The interrelation of DKK3 expression with the infiltration percentage of MDSCs in tumors from TCGA-CESC, ESCA, HNSC, MESO and SKCM

Article Snippet: They received intraperitoneal injections of 10 mg/kg of a mouse monoclonal DKK3 antibody (DKK3-4.22, BE0385, Bio X Cell), a mouse functional PD-1 antibody (BE0146, Bio X Cell), or a combination of both.

Techniques: Expressing

Journal: BMC cancer

Article Title: Targeting DKK3 to remodel tumor immune microenvironment and enhance cancer immunotherapy.

doi: 10.1186/s12885-025-14075-2

Figure Lengend Snippet: Fig. 3 DKK3 inhibits CD8+ T cell activation and Th1 differentiation ex vivo. (A) Primary CD4+ or CD8+ T cells isolated and sorted by MACS were treated with IL-2 (20 ng/mL) and IL-15 (20 ng/mL), and co-cultured with DKK3 recombinant protein (50 ng/mL). After 48 h co-culture, the activation level of CD8+ T cells was measured by the expressions of (B) CD25, (C) CD69, (D) CD107a and (E) IFN-γ by flow cytometry. And the expression of (F) T-bet and (G) GATA3 in CD4+ T cells were measured by flow cytometry. The unpaired student’s t-test is used to determine whether data with error bars are significant (*P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001)

Article Snippet: They received intraperitoneal injections of 10 mg/kg of a mouse monoclonal DKK3 antibody (DKK3-4.22, BE0385, Bio X Cell), a mouse functional PD-1 antibody (BE0146, Bio X Cell), or a combination of both.

Techniques: Activation Assay, Ex Vivo, Isolation, Cell Culture, Recombinant, Co-Culture Assay, Flow Cytometry, Expressing

Journal: BMC cancer

Article Title: Targeting DKK3 to remodel tumor immune microenvironment and enhance cancer immunotherapy.

doi: 10.1186/s12885-025-14075-2

Figure Lengend Snippet: Fig. 4 DKK3 blockade controls tumor growth in different mouse cancer models. (A-B) The tumor volume and overall survival of LLC-challenged mice with or without DKK3-4.22 treatment (10 mg/kg). (C-D) The tumor volume and overall survival of MC38-challenged mice with or without DKK3-4.22 treatment (10 mg/kg). (E-F) The tumor volume and overall survival of MFC-challenged mice with or without DKK3-4.22 treatment (10 mg/kg). (G-H) The tumor volume and overall survival of Pan02-challenged mice with or without DKK3-4.22 treatment (10 mg/kg). The unpaired student’s t-test is used to determine whether data with error bars are significant (*P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001)

Article Snippet: They received intraperitoneal injections of 10 mg/kg of a mouse monoclonal DKK3 antibody (DKK3-4.22, BE0385, Bio X Cell), a mouse functional PD-1 antibody (BE0146, Bio X Cell), or a combination of both.

Techniques:

Journal: BMC cancer

Article Title: Targeting DKK3 to remodel tumor immune microenvironment and enhance cancer immunotherapy.

doi: 10.1186/s12885-025-14075-2

Figure Lengend Snippet: Fig. 5 DKK3 blockade remodels the tumor immune microenvironment of different cancers. At the treatment endpoint, the percentages of (A) CD8+/ CD3+ cells, (B) IFNγ+/CD8+ T cells, (C) T-bet+/CD8+ cells, (D) CD164+/F4/80+ cells, and (E) Gr-1+/CD11b+ cells in tumors of the syngeneic LLC, MC38, MFC and Pan02 models were detected by flow cytometry (n = 5 per group). The unpaired student’s t-test is used to determine whether data with error bars are significant (*P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001)

Article Snippet: They received intraperitoneal injections of 10 mg/kg of a mouse monoclonal DKK3 antibody (DKK3-4.22, BE0385, Bio X Cell), a mouse functional PD-1 antibody (BE0146, Bio X Cell), or a combination of both.

Techniques: Flow Cytometry

Journal: BMC cancer

Article Title: Targeting DKK3 to remodel tumor immune microenvironment and enhance cancer immunotherapy.

doi: 10.1186/s12885-025-14075-2

Figure Lengend Snippet: Fig. 6 Combined blockade of DKK3 and PD-1 triggers synergistic anti-tumor effects. (A) The tumor volume of LLC-challenged mice with DKK3-4.22 (10 mg/kg) and / or anti-PD-1 treatment (10 mg/kg). (B) The tumor volume in each group of the LLC mouse model. The unpaired student’s t-test is used to determine whether data with error bars are significant (*P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001)

Article Snippet: They received intraperitoneal injections of 10 mg/kg of a mouse monoclonal DKK3 antibody (DKK3-4.22, BE0385, Bio X Cell), a mouse functional PD-1 antibody (BE0146, Bio X Cell), or a combination of both.

Techniques: