Journal: PloS one

Article Title: Wnt pathway activity in breast cancer sub-types and stem-like cells.

doi: 10.1371/journal.pone.0067811

Figure Lengend Snippet: Figure 3. Gene expression analysis of Wnt signalling in monolayer (Mono) and anoikis resistant (AR) cells of normal breast cell lines (N), ER-ve and ER+ve breast cancer cell lines. A) Protein expression of activated B-catenin (unphosphorylated), Lef1, Axin2, DKK1 and B-actin (housekeeper) in MCF7 monolayer and AR cells. B) Cluster analysis was performed using the fold change in expression

Article Snippet: MCF10a, MCF7, MDA-MB-231, primary human normal breast cells and primary human invasive breast cancer cells were plated into MS culture and treated with a single dose of human recombinant DKK1 (R and D systems) at increasing concentrations (0–100ng/ml).

Techniques: Gene Expression, Expressing

Journal: PloS one

Article Title: Wnt pathway activity in breast cancer sub-types and stem-like cells.

doi: 10.1371/journal.pone.0067811

Figure Lengend Snippet: Figure 4. Modulation of Wnt signalling in normal and breast cancer cell lines. Single cells were plated in non-adherent conditions and treated with increasing concentrations of either Wnt3a (0–50 ng/ml) or DKK1 (0–100 ng/ml) and cultured for 7 days and number of mammospheres counted. Wnt3a treatments are displayed in the left panel and DKK1 treatments in the right panel. Light grey bars represent untreated control A) MCF10a cells (Wnt3a) B) MCF7 cells (Wnt3a) C) MDA-MB-231 cells (Wnt3a) D) MCF10a cells (DKK1) E) MCF7 cells (DKK1) F) MDA-MB-231 cells (DKK1). Data is expressed as % mammosphere formation units. P values were generated by ANOVA. Asterisks mark individual comparisons which reached statistical significance * ,0.01 ** ,0.001 generated by a T-test. G) Image of a MCF10a mammosphere H) Image of an MCF7 mammosphere I) Image of an MDA-MB-231 mammosphere. Scale bar represents 50 mM. doi:10.1371/journal.pone.0067811.g004

Article Snippet: MCF10a, MCF7, MDA-MB-231, primary human normal breast cells and primary human invasive breast cancer cells were plated into MS culture and treated with a single dose of human recombinant DKK1 (R and D systems) at increasing concentrations (0–100ng/ml).

Techniques: Cell Culture, Control, Generated

Journal: PloS one

Article Title: Wnt pathway activity in breast cancer sub-types and stem-like cells.

doi: 10.1371/journal.pone.0067811

Figure Lengend Snippet: Figure 5. Modulation of Wnt signalling in normal and primary breast cancer samples (Normal n = 3; ER+ve n = 3; ER-ve n = 3). Single cells were plated in non-adherent conditions and treated with increasing concentrations of either Wnt3a (0–50 ng/ml) or DKK1 (0–100 ng/ml) and cultured for 7 days and number of mammospheres counted. Wnt3a treatments are displayed in the left panel and DKK1 treatments in the right panel. Light grey bars represent untreated control A) primary normal breast cells (Wnt3a) B) ER+ve primary breast cancer cells (Wnt3a) C) ER-ve primary breast cancer cells (Wnt3a) D) primary normal breast cells (DKK1) E) ER+ve primary breast cancer cells (DKK1) F) ER2ve primary breast cancer cells (DKK1). Data is expressed as % mammosphere formation units. P values were generated by ANOVA. Asterisks mark individual comparisons which reached statistical significance * .0.01 ** .0.001 generated by a T-test. G) Image of a normal primary mammosphere H) Image of an ER positive primary tumour mammosphere I) Image of an ER negative primary tumour mammosphere. Scale bar represents 50 mM. doi:10.1371/journal.pone.0067811.g005

Article Snippet: MCF10a, MCF7, MDA-MB-231, primary human normal breast cells and primary human invasive breast cancer cells were plated into MS culture and treated with a single dose of human recombinant DKK1 (R and D systems) at increasing concentrations (0–100ng/ml).

Techniques: Cell Culture, Control, Generated

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: eLife

Article Title: Genome-wide interrogation of extracellular vesicle biology using barcoded miRNAs

doi: 10.7554/eLife.41460

Figure Lengend Snippet: ( A ) Serum-starved K562 cells were treated with PBS (control) or Wnt3a (100 ng/ml) or Wnt3a + Dickkopf peptide (DKK1, 200 ng/ml) for 24 hr prior to EV isolation. Biochemical analysis of cellular (Cells) and EV fractions was performed by immunoblotting of extracts using antibodies against antigens shown. Detection of ß-Catenin was carried out using cytosolic fractions. ( B ) Quantitation of EV marker protein levels from EV immunoblots shown in ( A ). ( C ), Serum-starved K562 cells were treated with DMSO (control) or CHIR99021 (10 µM) for 24 hr prior to EV isolation. Biochemical analysis of EV fractions was performed as in ( A ). Representative immunoblots for a control experiment along with two replicate CHIR99021-treated samples are shown. ( D ) Quantitation of protein levels from EV immunoblots in ( C ). ( E ) Quantitation of LAMP1 protein from cellular fractions in ( C ). Molecular mass marker mobility is shown at the left of immunoblot panels in kilodaltons. t test: *p<0.05; **p<0.01; error bars represent SEM; n ≥ 3. ( F ) Flow cytometry determination of LAMP1 levels in anti-LAMP antibody-labeled control and CHIR99021-treated cells. Confocal images at right show lysosomes stained with anti-LAMP1 antibody (green) and nuclei labeled with DAPI (blue). Bar,10 µm.

Article Snippet: The next day, the medium was replaced with fresh EV-free RPMI supplemented with 100 ng/ml recombinant Wnt3a protein (R and D Systems), or Wnt3a combined with 200 ng/ml recombinant DKK1 protein (R and D Systems) and EVs were collected 24 hr later.

Techniques: Control, Isolation, Western Blot, Quantitation Assay, Marker, Flow Cytometry, Labeling, Staining

Journal: Oncogene

Article Title: A multi-faceted discovery strategy identifies functional antibodies binding to cysteine-rich domain 1 of hDKK1 for cancer immunotherapy via Wnt non-canonical pathway

doi: 10.1038/s41388-025-03445-6

Figure Lengend Snippet: A Two epitope bins are apparent amongst the Twist anti-DKK1 leads from this epitope binning analysis. The formation of Antibody-Antigen-Antibody complexes indicates the antibodies are not binding to the same epitope of DKK1. B Anti-DKK1 lead antibodies bind to hDKK1 cysteine-rich domain CRD1 or CRD2 or both CRD1 and CDR2 (in the instance of bispecific antibodies), and cross-reactivity with mouse and cynomolgus monkey DKK1. The assays were repeated in triplicate.

Article Snippet: On Day 2, mWNT3a (R&D Systems, #1324-WN, 50 ng/mL) and hDKK1 (R&D Systems, #5439-DK/CF, 500 ng/mL) were incubated with anti-DKK1 antibodies in assay buffer (BPS Bioscience Thaw medium 1, #60187 with 10 mM LiCl) at 37 °C for 30 min.

Techniques: Binding Assay

Journal: Oncogene

Article Title: A multi-faceted discovery strategy identifies functional antibodies binding to cysteine-rich domain 1 of hDKK1 for cancer immunotherapy via Wnt non-canonical pathway

doi: 10.1038/s41388-025-03445-6

Figure Lengend Snippet: A Wnt TCF/LEF reporter assay screening. Wnt TCF/LEF signaling is blocked by DKK1 binding to LRP5/6. Anti-DKK1 antibodies that bind to hDKK1 CRD2 block the binding of DKK1 to the co-receptors, and lead to the reactivation of Wnt canonical signaling. B MC3T3.E1 cell differentiation detection by mineralization assay. Soluble hDKK1 suppresses pre-osteoblast cell differentiation via the Wnt canonical pathway. Anti-DKK1 antibodies that bind to hDKK1 CRD2 block the binding of DKK1 to the LRP5/6 co-receptors and restore cell differentiation. C Wnt non-canonical phospho-JNK detection. Colo205 cells were treated with Wnt, DKK1, and anti-DKK1 lead antibodies. Intracellular JNK phosphorylation level was detected with ELISA. D Primary immune cell activation. DKK1 leads to immune suppression including T cell inactivation, MDSC accumulation, and NK cell clearance. GM-CSF is the biomarker for NK cell activation. Human PBMC were treated with an immune stimulator, mWNT3a, hDKK1, and DKK1 lead antibodies. Cytokine release of GM-CSF was measured by ELISA. Antibodies binding to CRD1 of DKK1 showed stronger NK cell activation. E PC3 tumor cell cytotoxicity by activated immune cells. T cells and NK cells in human PBMC were activated and co-cultured with PC3 tumor cells for 6 days. Activated immune cells kill PC3 cells, while hDKK1 treatment inhibits cytotoxicity. Blocking the interaction of hDKK1 to the receptor with Twist DKK1 lead antibodies restores the cytotoxicity potency. Antibodies binding to CRD1 of DKK1 showed stronger cytotoxicity. F Anti-DKK1 antibody targeting DKK1 CRD1 also induced cytotoxicity in breast, gastric and colon cancer cells. Using the KILR cytotoxicity assay, a high luminescence signal was detected in cytotoxic cells. All the samples were duplicated, and the assays were repeated three times.

Article Snippet: On Day 2, mWNT3a (R&D Systems, #1324-WN, 50 ng/mL) and hDKK1 (R&D Systems, #5439-DK/CF, 500 ng/mL) were incubated with anti-DKK1 antibodies in assay buffer (BPS Bioscience Thaw medium 1, #60187 with 10 mM LiCl) at 37 °C for 30 min.

Techniques: Reporter Assay, Binding Assay, Blocking Assay, Cell Differentiation, Mineralization Assay, Phospho-proteomics, Enzyme-linked Immunosorbent Assay, Activation Assay, Biomarker Discovery, Cell Culture, Cytotoxicity Assay

Journal: Journal of personalized medicine

Article Title: Impact of REAC Regenerative Endogenous Bioelectrical Cell Reprogramming on MCF7 Breast Cancer Cells.

doi: 10.3390/jpm13061019

Figure Lengend Snippet: Figure 7. DKK1 and SFRP1 quantification by ELISA. The concentrations of DKK1 (A) and SFRP1 (B) were measured after 7, 10, and 14 days in supernatants of MCF7 either exposed or not to REAC TO-RGN treatment. Data are expressed as mean ± SD relative to the control (mean ± SD) (** p ≤0.01; *** p ≤0.001).

Article Snippet: The concentrations of human secreted frizzled-related protein 1 (SFRP1) and human Dickkopf-related protein 1 (DKK1) were determined using the human secreted frizzledrelated protein 1 (SFRP1) ELISA Kit (Cusabio, Flarebio Biotech LLC) and the human DKK1 (Dickkopf-related protein 1) ELISA Kit (Elabscience Biotechnology Co., Ltd, Houston, TX, USA), respectively.

Techniques: Enzyme-linked Immunosorbent Assay, Control

Journal: Frontiers in oncology

Article Title: Chondroitin Sulfates Control Invasiveness of the Basal-Like Breast Cancer Cell Line MDA-MB-231 Through ROR1.

doi: 10.3389/fonc.2022.914838

Figure Lengend Snippet: FIGURE 6 | CS-E-elicited invasiveness is enhanced by the absence of DKK1. (A) Raw sensor grams. ROR1 was immobilized in a flow cell of a CM5 sensor chip. DKK1 alone (a), DKK1 premixed with CS-E at a 1:7 molar ratio (b), and DKK1 premixed with CS-A at a 1:7 molar ration (c) were used as analytes. (B) Response- unit quantification of binding. (C) DKK1 mRNA expression in MDA-MB-231 cells transfected with siDKK1 or control siRNA (siCont) measured using qPCR (n=4). Expression data were normalized to those of GAPDH. (D) Invasiveness of DKK1 knocked down MDA-MB-231 cells (siDKK1) or control cells (siCont) treated with or without CS-E (n>5). Data were analyzed using a Tukey–Kramer multiple comparison.

Article Snippet: For ROR1 binding assays, WNT5A (0, 0.038, 0.075, 0.15, 0.30, and 0.60 mM), WNT5A/CS-E (0, 0.036/0.25, 0.071/0.50, 0.15/1.0, 0.29/ 2.0, and 0.57/4.0 mM), recombinant human DKK1 (Cat. No. 5439-DK/CF, R&D Systems) (0, 0.038, 0.075, 0.15, and 0.3 mM), or DKK1/CS-E (0, 0.038/0.28, 0.075/0.55, 0.15/1.1, and 0.3/2.1 mM) were sequentially injected at a flowrate of 30 ml/ min for 120 s at 25°C; the dissociation time was set for 130 s. Binding reactions were performed in 50 mM Tris–HCl buffer (pH 7.5).

Techniques: Binding Assay, Expressing, Transfection, Control, Comparison

Journal: Frontiers in oncology

Article Title: Chondroitin Sulfates Control Invasiveness of the Basal-Like Breast Cancer Cell Line MDA-MB-231 Through ROR1.

doi: 10.3389/fonc.2022.914838

Figure Lengend Snippet: FIGURE 7 | Schematic of CS-E enhancement of invasive activity of the triple-negative breast cancer MDA-MB-231 cell line. CS chains bind WNT5A and ROR1 through E units, signaling cancer cells to activate JNK1. Decreasing E units by knockdown of CHST11 and CHST15 inhibits WNT5A−ROR1−JNK signaling. DKK1 suppresses CS tumor promoting activity by binding to E units.

Article Snippet: For ROR1 binding assays, WNT5A (0, 0.038, 0.075, 0.15, 0.30, and 0.60 mM), WNT5A/CS-E (0, 0.036/0.25, 0.071/0.50, 0.15/1.0, 0.29/ 2.0, and 0.57/4.0 mM), recombinant human DKK1 (Cat. No. 5439-DK/CF, R&D Systems) (0, 0.038, 0.075, 0.15, and 0.3 mM), or DKK1/CS-E (0, 0.038/0.28, 0.075/0.55, 0.15/1.1, and 0.3/2.1 mM) were sequentially injected at a flowrate of 30 ml/ min for 120 s at 25°C; the dissociation time was set for 130 s. Binding reactions were performed in 50 mM Tris–HCl buffer (pH 7.5).

Techniques: Activity Assay, Knockdown, Binding Assay

Journal: International Journal of Medical Sciences

Article Title: LRP5 enhances glioma cell proliferation by modulating the MAPK/p53/cdc2 pathway

doi: 10.7150/ijms.99920

Figure Lengend Snippet: The LRP5 antagonist DKK-1 reduced the proliferation of glioma cells through the modulation of MAPK/p53/cdc2 pathway. (A-C) Representative images and the statistical results of Edu assay in A172 and SHG-44 cells (n=3). The nucleus was stained with Hoechst (blue), and the proliferating cells were stained with Edu (red). (scale bar=250μm) ** P <0.01, *** P <0.001. (D-G) Representative images of protein levels for p53, p-p53, cdc2, p-cdc2, JNK, p-JNK, p38, and p-p38 were analyzed by Western blotting. (H, I) The statistical results of Western blotting in A172 and SHG-44 cells treated with DKK-1(0.5μg/mL) or a negative control for 48h. ns P 0.05, * P <0.05, ** P <0.01, *** P 0.001. All data were presented as the means ± SD.

Article Snippet: DKK-1 was obtained from MedChemExpress (MCE, HY-P72968).

Techniques: EdU Assay, Staining, Western Blot, Negative Control

Journal: Oncology Letters

Article Title: Role of prognostic gene DKK1 in oral squamous cell carcinoma

doi: 10.3892/ol.2023.14184

Figure Lengend Snippet: Expression of DKK1 in various tumor types. Expression levels of DKK1 in (A) The Cancer Genome Atlas, (B) GSE3524 and (C) GSE37991 oral squamous cell carcinoma datasets. (D) Expression level of DKK1 in 33 cancer types. *P<0.05, **P<0.01, ***P<0.001. DKK1, Dickkopf Wnt signaling pathway inhibitor 1; TPM, transcripts per million; ns, not significant; ACC, adrenocortical carcinoma; BLCA, bladder urothelial carcinoma; BRCA, breast invasive carcinoma; CESC, cervical squamous cell carcinoma and endocervical adenocarcinoma; CHOL, cholangiocarcinoma; COAD, colon adenocarcinoma; DLBC, lymphoid neoplasm diffuse large B-cell lymphoma; ESCA, esophageal carcinoma; GBM, glioblastoma multiforme; HNSC, head and neck squamous cell carcinoma; KICH, kidney chromophobe; KIRC, kidney renal clear cell carcinoma; KIRP, kidney renal papillary cell carcinoma; LAML, acute myeloid leukemia; LGG, brain lower grade glioma; LIHC, liver hepatocellular carcinoma; LUAD, lung adenocarcinoma; LUSC, lung squamous cell carcinoma; MESO, mesothelioma; OV, ovarian serous cystadenocarcinoma; PAAD, pancreatic adenocarcinoma; PCPG, pheochromocytoma and paraganglioma; PRAD, prostate adenocarcinoma; READ, rectum adenocarcinoma; SARC, sarcoma; SKCM, skin cutaneous melanoma; STAD, stomach adenocarcinoma; TGCT, testicular germ cell tumors; THCA, thyroid carcinoma; THYM, thymoma; UCEC, uterine corpus endometrial carcinoma; UCS, uterine carcinosarcoma; UVM, uveal melanoma.

Article Snippet: Secreted DKK1 in the culture supernatant was detected using a Human DKK1 ELISA Kit (cat. no. EK0867; Wuhan Boster Biological Technology, Ltd.) following the manufacturer's instructions.

Techniques: Expressing

Journal: Oncology Letters

Article Title: Role of prognostic gene DKK1 in oral squamous cell carcinoma

doi: 10.3892/ol.2023.14184

Figure Lengend Snippet: Univariate survival analysis of the association between DKK1 expression and survival time in 12 tumor types. (A) Forest plot showing the relationship between DKK1 expression and OS. KM curves of high and low DKK1 expression in (B) ESCA, (C) HNSC and (D) STAD reveal a significant association with OS. (E) Forest plot showing the relationship between DKK1 expression and DSS. KM curves of high and low DKK1 expression in (F) HNSC and (G) STAD show a significant association with DSS. DKK1, Dickkopf Wnt signaling pathway inhibitor 1; OS, overall survival; DSS, disease-specific survival; KM, Kaplan-Meier; BLCA, bladder urothelial carcinoma; CHOL, cholangiocarcinoma; COAD, colon adenocarcinoma; ESCA, esophageal carcinoma; HNSC, head and neck squamous cell carcinoma; KICH, kidney chromophobe; KIRP, kidney renal papillary cell carcinoma; LIHC, liver hepatocellular carcinoma; LUSC, lung squamous cell carcinoma; PRAD, prostate adenocarcinoma; STAD, stomach adenocarcinoma; THCA, thyroid carcinoma; HR, hazard ratio.

Article Snippet: Secreted DKK1 in the culture supernatant was detected using a Human DKK1 ELISA Kit (cat. no. EK0867; Wuhan Boster Biological Technology, Ltd.) following the manufacturer's instructions.

Techniques: Expressing

Journal: Oncology Letters

Article Title: Role of prognostic gene DKK1 in oral squamous cell carcinoma

doi: 10.3892/ol.2023.14184

Figure Lengend Snippet: Correlation analysis of DKK1 and immune cell infiltration. (A) Correlation of DKK1 with the infiltration of a panel of immune cells. Negative correlation of DKK1 expression with the level of infiltration of (B) T cells, (C) Treg cells, (D) pDC cells, (E) Th17 cells, (F) TFH cells, (G) cytotoxic cells and (H) B cells. Positive correlation of DKK1 expression with the level of infiltration of (I) Th2 cells and (J) Tgd cells. DKK1, Dickkopf Wnt signaling pathway inhibitor 1; DC, dendritic cell; aDC, activated DC; iDC, inflammatory DC; pDC, plasmacytoid DC; NK, natural killer; Tcm, T central memory; Tem, T effector memory; TFH, T follicular helper; Tgd, γδT; Th, T helper; TReg, regulatory T; TPM, transcripts per million.

Article Snippet: Secreted DKK1 in the culture supernatant was detected using a Human DKK1 ELISA Kit (cat. no. EK0867; Wuhan Boster Biological Technology, Ltd.) following the manufacturer's instructions.

Techniques: Expressing

Journal: Oncology Letters

Article Title: Role of prognostic gene DKK1 in oral squamous cell carcinoma

doi: 10.3892/ol.2023.14184

Figure Lengend Snippet: Correlation between DKK1 and genes associated with immune checkpoints in oral squamous cell carcinoma. DKK1 expression was positively correlated with the expression levels of (A) CD40, (B) CD44, (C) VTCN1, (D) NRP1, (E) PDCD1LG2, (F) CD276, (G) CD80 and (H) CD86 and negatively correlated with expression levels of (I) TNFRSR18, (J) CD27, (K) TIGIT, (L) IDO2, (M) CD48, (N) CD244 and (O) CD40LG. DKK1, Dickkopf Wnt signaling pathway inhibitor 1; VTCN1, v-set domain containing T cell activation inhibitor 1; NRP1, neuropilin 1; PDCD1LG2, programmed cell death 1 ligand 2; TNFRSR18, TNF receptor superfamily member 18; TIGIT, T cell immunoreceptor with Ig and ITIM domains; IDO2, indoleamine 2,3-dioxygenase 2; CD40LG, CD40 ligand; TPM, transcripts per million.

Article Snippet: Secreted DKK1 in the culture supernatant was detected using a Human DKK1 ELISA Kit (cat. no. EK0867; Wuhan Boster Biological Technology, Ltd.) following the manufacturer's instructions.

Techniques: Expressing, Activation Assay

Journal: Oncology Letters

Article Title: Role of prognostic gene DKK1 in oral squamous cell carcinoma

doi: 10.3892/ol.2023.14184

Figure Lengend Snippet: Correlation of DKK1 expression with that of five DNA mismatch repair genes in oral squamous cell carcinoma. *P<0.05; **P<0.01; ***P<0.001. DKK1, Dickkopf Wnt signaling pathway inhibitor 1; MLH1, MutL homolog 1; MSH2/6, MutS homolog 2/6; PMS2, PMS1 homolog 2; EPCAM, epithelial cell adhesion molecule.

Article Snippet: Secreted DKK1 in the culture supernatant was detected using a Human DKK1 ELISA Kit (cat. no. EK0867; Wuhan Boster Biological Technology, Ltd.) following the manufacturer's instructions.

Techniques: Expressing

Journal: Oncology Letters

Article Title: Role of prognostic gene DKK1 in oral squamous cell carcinoma

doi: 10.3892/ol.2023.14184

Figure Lengend Snippet: Knockdown of DKK1 inhibits cell proliferation, colony formation, migration and invasion in oral squamous cell carcinoma cells. Bright-field and GFP fluorescence images of (A) SCC-9 cells and (B) CAL-27 cells with Lv-shCon and Lv-shDKK1 infection. Scale Bar=50 µm. (C) DKK1 mRNA expression and (D) DKK1 secretion by SCC-9 cells and CAL-27 cells infected with Lv-shCon and Lv-shDKK1. Cell proliferation rate of (E) SCC-9 and (F) CAL-27 cells. (G) Colony formation of SCC-9 cells and CAL-27 cells. (H) Representative wound healing images of SCC-9 cells and (I) quantification of the wound healing assay. (J) Representative wound healing images of CAL-27 cells and (K) quantification of the wound healing assay, magnification: 40×. Representative images of the Transwell invasion assay for (L) SCC-9 and (M) CAL-27 cells, magnification: 100×. (N) Cell counts of invaded SCC-9 and CAL-27 cells. (O) Western blot analysis of Wnt-3a, β-catenin and DKK1 expression in SCC-9 and CAL-27 cells infected with Lv-shCon and Lv-shDKK1. *P<0.05 **P<0.01 and ***P<0.001 for shDKK1 vs. shCon. DKK1, Dickkopf Wnt signaling pathway inhibitor 1; GFP, green fluorescent protein; Lv, lentivirus; shCon, shRNA control; shDKK1, shRNA targeting DKK1; shRNA, short hairpin RNA.

Article Snippet: Secreted DKK1 in the culture supernatant was detected using a Human DKK1 ELISA Kit (cat. no. EK0867; Wuhan Boster Biological Technology, Ltd.) following the manufacturer's instructions.

Techniques: Knockdown, Migration, Fluorescence, Infection, Expressing, Wound Healing Assay, Transwell Invasion Assay, Western Blot, shRNA, Control