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: 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: Life Science Alliance

Article Title: Hepatic DKK1-driven steatosis is CD36 dependent

doi: 10.26508/lsa.202201665

Figure Lengend Snippet: (A) Immunohistochemistry staining of DKK1 in the livers of patients with NAFLD (n = 5) and normal individuals (n = 3); scale bar = 100 µ m, (B) in liver samples from WT mice fed with chow and HFD for 24 wk (n = 5). Scale bar = 100 µ m. (C) qPCR analysis of DKK1 mRNA expression in livers of chow- and HFD-fed 24-wk mice (n = 4 mice for each group). (D, E) Representative Western blot of DKK1 in liver samples of chow- and HFD-fed mice (n = 4). Each lane represents liver lysates from individual mouse. (F) Serum DKK1 protein levels in chow- or HFD-fed mice for 24 wk (n = 15–17). (G) Western blot of DKK1 in different primary cells isolated from HFD-fed mice liver. Hep, hepatocytes; HSC, hepatic stellate cells; LSEC, liver sinusoidal endothelial cells; KC, Kupffer cells. (H) Western blot analysis of DKK1 in AML12 cells under different concentrations of FFA exposures (N = 2). ** P < 0.01 as compared with the indicated controls by two-tailed t tests (two groups). All data are shown as the means ± SD. Source data are available for this figure.

Article Snippet: ALT, AST, TC, TG levels were measured using an automatic biochemical analyzer (7020; Hitachi) in Fengrui Biotechnology Co. Serum levels of DKK1 were measured using a commercially available mouse ELISA kit (EK0925; Boster Biological) according to the manufacturer’s instructions.

Techniques: Immunohistochemistry, Staining, Expressing, Western Blot, Isolation, Two Tailed Test

Journal: Life Science Alliance

Article Title: Hepatic DKK1-driven steatosis is CD36 dependent

doi: 10.26508/lsa.202201665

Figure Lengend Snippet: (A) Represented images of DKK1 immunohistochemistry staining on liver sections from NAFLD patients (n = 5) and normal controls (n = 3); scale bar = 100 µ m. (B) LW and LW/BW index of chow- and HFD-fed 6-mo mice (n = 5). (C) Lipid accumulations in the livers were analyzed by H&E staining and ORO staining (n = 3); scale bar = 100 µ m. (D, E) Serum ALT, AST; (E) serum lipids measurement (n = 14–17). ** P < 0.01, *** P < 0.001, **** P < 0.0001 as compared with the indicated controls by two-tailed t tests. All data are shown as the means ± SD. Source data are available for this figure.

Article Snippet: ALT, AST, TC, TG levels were measured using an automatic biochemical analyzer (7020; Hitachi) in Fengrui Biotechnology Co. Serum levels of DKK1 were measured using a commercially available mouse ELISA kit (EK0925; Boster Biological) according to the manufacturer’s instructions.

Techniques: Immunohistochemistry, Staining, Two Tailed Test

Journal: Life Science Alliance

Article Title: Hepatic DKK1-driven steatosis is CD36 dependent

doi: 10.26508/lsa.202201665

Figure Lengend Snippet: (A) Schematic illustration of experiment procedure. AAV tail intravenous injection with AAV-GFP-NC (n = 4), AAV-OE-DKK1 (n = 5), or AAV-sh-DKK1 (n = 6) and then fed HFD for 20 wk before euthanasia. (B) Dynamic body weight tracking of chow- and HFD-fed mice with DKK1 manipulations. (C) Representative Western blot of DKK1 in liver samples of HFD-fed mice after 20 wk (n = 3). Each lane represents liver lysates from individual mouse. (D) H&E and ORO staining. Scale bar = 200 µ m; (E, F) lipid contents in liver of mice with different DKK1 gene manipulations under chow or HFD fed (n = 3). (G, H) Serum lipid contents with different DKK1 gene manipulations under chow or HFD fed. (I) The expression confirmations of lipid metabolism–related genes in liver of mice with different DKK1 gene manipulations under chow or HFD fed (n = 3). * P < 0.05, ** P < 0.01, *** P < 0.001 as compared with the indicated controls by two-tailed t tests. All data are shown as the means ± SD. ns, not significant. Source data are available for this figure.

Article Snippet: ALT, AST, TC, TG levels were measured using an automatic biochemical analyzer (7020; Hitachi) in Fengrui Biotechnology Co. Serum levels of DKK1 were measured using a commercially available mouse ELISA kit (EK0925; Boster Biological) according to the manufacturer’s instructions.

Techniques: Injection, Western Blot, Staining, Expressing, Two Tailed Test

Journal: Life Science Alliance

Article Title: Hepatic DKK1-driven steatosis is CD36 dependent

doi: 10.26508/lsa.202201665

Figure Lengend Snippet: (A) H&E staining of livers from controls and different DKK1 gene manipulations with 20 wk chow or HFD fed; scale bar = 100 µ m. (B) mRNA levels of Wnt-related genes in WT and DKK1-KO HepG2 cells treated with FFA or BSA for 24 h (n = 3). (C) Representative Western blot of ROCK1, total and phosphorylated JNK in WT, and DKK1-KO HepG2 cells treated with FFA or BSA for 24 h (N = 2). * P < 0.05, ** P < 0.01 as compared with the indicated controls by two-tailed t tests. All data are shown as the means ± SD. Source data are available for this figure.

Article Snippet: ALT, AST, TC, TG levels were measured using an automatic biochemical analyzer (7020; Hitachi) in Fengrui Biotechnology Co. Serum levels of DKK1 were measured using a commercially available mouse ELISA kit (EK0925; Boster Biological) according to the manufacturer’s instructions.

Techniques: Staining, Western Blot, Two Tailed Test

Journal: Life Science Alliance

Article Title: Hepatic DKK1-driven steatosis is CD36 dependent

doi: 10.26508/lsa.202201665

Figure Lengend Snippet: (A, B, C, D, E, F, G, H) The targeted locus of DKK1 and sequencing analysis of homozygous knockout HepG2 and AML12 (A, B) cells that were confirmed with Western blot (C, D), and the paralleled experiments performed on CD36 knockout cells (E, F, G, H). (I, J, K) The DKK1-overexpressed (OE-DKK1) or DKK1-knock-down cell lines (sh-DKK1) were constructed by transfecting overexpression lentivirus or shRNA lentivirus, respectively. Scale bar = 200 µ m (I), and the DKK1 levels were confirmed by Western blot analysis (J, K). Source data are available for this figure.

Article Snippet: ALT, AST, TC, TG levels were measured using an automatic biochemical analyzer (7020; Hitachi) in Fengrui Biotechnology Co. Serum levels of DKK1 were measured using a commercially available mouse ELISA kit (EK0925; Boster Biological) according to the manufacturer’s instructions.

Techniques: Sequencing, Knock-Out, Western Blot, Knockdown, Construct, Over Expression, shRNA

Journal: Life Science Alliance

Article Title: Hepatic DKK1-driven steatosis is CD36 dependent

doi: 10.26508/lsa.202201665

Figure Lengend Snippet: (A, B, C) Under DKK1 knockout conditions, cell steatosis was induced by FFA (A, B, C), n = 3 in each group. (A, B, C) Oil red O staining (A) scale bar = 50 µ m; TG measurements in DKK1 −/− AML12 (B) and DKK1 −/− HepG2 (C) cells. n = 3 in each group. (D, E) The changed TG was further confirmed with administration of a DKK1 inhibitor, WAY262611 in AML12 (D) and HepG2 (E) cells. n = 3 in each group. (F, G, H) On other hand, under DKK1 overexpression condition, the induced steatosis status was parallel analyzed, oil red O staining (F), and TG measurements in DKK1 −/− AML12 (G) and DKK1 −/− HepG2 (H) cells. n = 3 in each group. (I, J) Furthermore, the changed trend of TG was further confirmed with administration of recombinant DKK1 protein in AML12 (I) and HepG2 (J) cells. n = 3 in each group. * P < 0.05, ** P < 0.01, *** P < 0.001 as compared with the indicated controls by two-tailed t tests. All data are shown as the means ± SD. Source data are available for this figure.

Article Snippet: ALT, AST, TC, TG levels were measured using an automatic biochemical analyzer (7020; Hitachi) in Fengrui Biotechnology Co. Serum levels of DKK1 were measured using a commercially available mouse ELISA kit (EK0925; Boster Biological) according to the manufacturer’s instructions.

Techniques: Knock-Out, Staining, Over Expression, Recombinant, Two Tailed Test

Journal: Life Science Alliance

Article Title: Hepatic DKK1-driven steatosis is CD36 dependent

doi: 10.26508/lsa.202201665

Figure Lengend Snippet: (A, B) Volcano map (A) and KEGG analysis (B) of up- and down-regulated genes in LV-OE-DKK1 AML12 cells compared with LV-GFP-NC AML12 cells under FFA exposure for 24 h. (C, D, E, F) The expression confirmations of lipid metabolism–related genes in cell lines with different DKK1 gene manipulations under FFA treatment; DKK1 overexpression in AML12 (C) and HepG2 (D); DKK1 knockout AML12 (E) and HepG2 (F) cells. n = 3 in each group. * P < 0.05, ** P < 0.01, *** P < 0.001 as compared with the indicated controls by two-tailed t tests. All data are shown as the means ± SD. ns, not significant. Source data are available for this figure.

Article Snippet: ALT, AST, TC, TG levels were measured using an automatic biochemical analyzer (7020; Hitachi) in Fengrui Biotechnology Co. Serum levels of DKK1 were measured using a commercially available mouse ELISA kit (EK0925; Boster Biological) according to the manufacturer’s instructions.

Techniques: Expressing, Over Expression, Knock-Out, Two Tailed Test

Journal: Life Science Alliance

Article Title: Hepatic DKK1-driven steatosis is CD36 dependent

doi: 10.26508/lsa.202201665

Figure Lengend Snippet: (A, B) Representative Western blot analyses of lipid metabolism–related proteins in DKK1 overexpression- or knock-down mice (A) and DKK1 overexpression in AML12 cell line (B), in which the numbers marked above the controls were the ratio between CD36 compared with the β-Tubulin. (C, D, E, F) The ORO staining of WT and CD36 −/− cells cultured with recombinant DKK1 protein (r-hDKK1 or r-mDKK1) with or without FFA induction (C, D) scale bar = 100 µ m, and measurement of TG contents in CD36 −/− HepG2 (E) and CD36 −/− AML12 (F) cells. n = 3 in each group. (G) The CD36-promoter-driven luciferase reporter assay under DKK1 overexpression condition. n = 3 in each group. (H) Western blot analysis shows the overexpression of DKK1 increased pERK and nucleus PPARγ in AML12 cells with independent duplicates. * P < 0.05, ** P < 0.01, *** P < 0.001 as compared with the indicated controls by two-tailed t tests. All data are shown as the means ± SD. Source data are available for this figure.

Article Snippet: ALT, AST, TC, TG levels were measured using an automatic biochemical analyzer (7020; Hitachi) in Fengrui Biotechnology Co. Serum levels of DKK1 were measured using a commercially available mouse ELISA kit (EK0925; Boster Biological) according to the manufacturer’s instructions.

Techniques: Western Blot, Over Expression, Knockdown, Staining, Cell Culture, Recombinant, Luciferase, Reporter Assay, Two Tailed Test

Journal: Life Science Alliance

Article Title: Hepatic DKK1-driven steatosis is CD36 dependent

doi: 10.26508/lsa.202201665

Figure Lengend Snippet: (A, B) The GTT (A) and ITT (B) analyses of mice with treatments of AAV-GFP-NC, AAV-OE-DKK1, or AAV-sh-DKK1 and HFD-fed 20 wk (n = 3). (C) Western blot analyses of phosphorylated JNK in AML12 cells under rDKK1 stimulation (N = 2). (D) Representative Western blot analyses of phosphorylated AKT and FOXO1 in AML12 cells under insulin and rDKK1 stimulation (N = 2). * P < 0.05, ** P < 0.01, *** P < 0.001 as compared with the indicated controls by two-tailed t tests. All data are shown as the means ± SD. Source data are available for this figure.

Article Snippet: ALT, AST, TC, TG levels were measured using an automatic biochemical analyzer (7020; Hitachi) in Fengrui Biotechnology Co. Serum levels of DKK1 were measured using a commercially available mouse ELISA kit (EK0925; Boster Biological) according to the manufacturer’s instructions.

Techniques: Western Blot, Two Tailed Test

Journal: Life Science Alliance

Article Title: Hepatic DKK1-driven steatosis is CD36 dependent

doi: 10.26508/lsa.202201665

Figure Lengend Snippet: In response to continuous challenge with HFD, (1) the DKK1 expression is mainly induced in hepatocytes and the increased serum DKK1 could have served as a diagnostic bio-marker for steatohepatitis progression. (2) DKK1 enhances hepatic CD36 expression by activating ERK-PPARγ signaling and consequently leads to increased hepatic fatty acid uptake and hepatocyte steatosis. (3) DKK1 activates JNK to decrease the phosphorylation of AKT and FOXO1, leading to insulin resistance. Hepatic fatty acid uptake and insulin resistance synergistically exacerbate fatty acid accumulation and the resultant hepatic steatosis.

Article Snippet: ALT, AST, TC, TG levels were measured using an automatic biochemical analyzer (7020; Hitachi) in Fengrui Biotechnology Co. Serum levels of DKK1 were measured using a commercially available mouse ELISA kit (EK0925; Boster Biological) according to the manufacturer’s instructions.

Techniques: Expressing, Diagnostic Assay, Marker, Phospho-proteomics

Journal: Life Science Alliance

Article Title: Hepatic DKK1-driven steatosis is CD36 dependent

doi: 10.26508/lsa.202201665

Figure Lengend Snippet: List of primers used for DKK1 and CD36 knock out and identify.

Article Snippet: ALT, AST, TC, TG levels were measured using an automatic biochemical analyzer (7020; Hitachi) in Fengrui Biotechnology Co. Serum levels of DKK1 were measured using a commercially available mouse ELISA kit (EK0925; Boster Biological) according to the manufacturer’s instructions.

Techniques: Knock-Out, Sequencing

Journal: Life Science Alliance

Article Title: Hepatic DKK1-driven steatosis is CD36 dependent

doi: 10.26508/lsa.202201665

Figure Lengend Snippet: Antibodies used for Western blot and IHC.

Article Snippet: ALT, AST, TC, TG levels were measured using an automatic biochemical analyzer (7020; Hitachi) in Fengrui Biotechnology Co. Serum levels of DKK1 were measured using a commercially available mouse ELISA kit (EK0925; Boster Biological) according to the manufacturer’s instructions.

Techniques: Western Blot