Journal: Cell reports

Article Title: ΔN-Tp63 Mediates Wnt/β-Catenin-Induced Inhibition of Differentiation in Basal Stem Cells of Mucociliary Epithelia

doi: 10.1016/j.celrep.2019.08.063

Figure Lengend Snippet: (A–F) BCIs in ALI culture for up to 4 weeks. Human recombinant DKK1 (DKK1) was used to inhibit Wnt/β-catenin signaling starting at ALI day 7 (D7). (A) Confocal imaging of specimens stained for Ac.-α-tubulin (MCCs, blue), CC10 (Club cells, green), and Actin (cell membranes, magenta) shows moderately increased MCC and Club cell numbers after DKK1 treatment. (B) DKK1 leads to a transient decrease in BCs but does not lead to loss of ΔN-TP63+ (green) cells. Nuclei (DAPI, blue). (C) Quantification from (A) and (B), respectively. Mann Whitney test, not significant, ns (p > 0.05); ***p ≤ 0.001. (D) Quantitative real-time PCR expression levels are depicted relative to stage controls. DKK1 increases expression of MCC ( FOXJ1 , MCIDAS ) and to a lesser extent Club cell ( SCGB1A1 ) markers but without reduction of BC markers ( ΔN-TP63 , KRT5 ). Student’s t test, not significant, ns (p > 0.05); *p ≤ 0.05; **p ≤ 0.01. (E and F) Optical orthogonal sections of confocal images after staining for BC markers ΔN-TP63 (green, in E) and KRT5 (green, in F). Related to .

Article Snippet: Manipulations of Wnt signaling were done by addition of human recombinant RSPO2 (R&D systems 3266-RS) or human recombinant DKK1 (R&D systems 5439-DK), which were reconstituted in sterile PBS, pH 7.4 containing 0.1% bovine serum albumin at 200ng/ml.

Techniques: Recombinant, Imaging, Staining, MANN-WHITNEY, Real-time Polymerase Chain Reaction, Expressing

Journal: Cell reports

Article Title: ΔN-Tp63 Mediates Wnt/β-Catenin-Induced Inhibition of Differentiation in Basal Stem Cells of Mucociliary Epithelia

doi: 10.1016/j.celrep.2019.08.063

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Manipulations of Wnt signaling were done by addition of human recombinant RSPO2 (R&D systems 3266-RS) or human recombinant DKK1 (R&D systems 5439-DK), which were reconstituted in sterile PBS, pH 7.4 containing 0.1% bovine serum albumin at 200ng/ml.

Techniques: Recombinant, RNA Library Preparation, cDNA Synthesis, SYBR Green Assay, Quantitative RT-PCR, Software

Journal: New England Journal of Medicine

Article Title: The Role of the Wnt-Signaling Antagonist DKK1 in the Development of Osteolytic Lesions in Multiple Myeloma

doi: 10.1056/nejmoa030847

Figure Lengend Snippet: Figure 2. Overexpression of DKK1 in Low-Grade Myeloma, with the Loss of Expression with Disease Progression.

Article Snippet: Serial dilutions of recombinant human DKK1 (R&D Systems) were used to establish a standard curve.

Techniques: Over Expression, Expressing, Biomarker Discovery

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: 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 μM), 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 μM), recombinant human DKK1 (Cat. No. 5439-DK/CF, R&D Systems) (0, 0.038, 0.075, 0.15, and 0.3 μM), or DKK1/CS-E (0, 0.038/0.28, 0.075/0.55, 0.15/1.1, and 0.3/2.1 μM) were sequentially injected at a flowrate of 30 μl/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: 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 μM), 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 μM), recombinant human DKK1 (Cat. No. 5439-DK/CF, R&D Systems) (0, 0.038, 0.075, 0.15, and 0.3 μM), or DKK1/CS-E (0, 0.038/0.28, 0.075/0.55, 0.15/1.1, and 0.3/2.1 μM) were sequentially injected at a flowrate of 30 μl/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: iScience

Article Title: Platelet DKK1 promotes tolerogenic dendritic cells and non-healing responses in cutaneous leishmaniasis

doi: 10.1016/j.isci.2026.115090

Figure Lengend Snippet: Decreased MPO + , CD11b + , and MHC class II + neutrophils in MyD88 (PKO) - and DKK1 (PKO) -infected mice (A–G) Six-week-old female WT-BALB/c, MyD88 (PKO) and DKK1 (PKO) mice were challenged with infective metacyclic promastigote (2 ×10 6 parasites, n = 5 per group) of L . major via the footpad. Non-infected BALB/c mice ( n = 10/2 feet per mouse) were given 0.9% NaCl saline. Cells were isolated from the footpads of all infected and non-infected mice at days 3 and 14 PI. Samples were analyzed by flow cytometry for MPO + , CD11b + , and MHC class II + neutrophils. A representative flow cytometry dot plot of MPO + , CD11b + , and MHC class II + neutrophils on day 3 is presented in . The percentage of MPO + , CD11b + , and MHC class II + cells in the different experimental groups is shown in column graphs (A, B, C, D, E, and F), while the absolute number of neutrophils obtained on day 3 PI is indicated (G). In all experiments, infected and non-infected BALB/c mice served as positive and negative controls, respectively. Results are presented as mean (±SEM) and are representative of two independent experiments. One-way ANOVA followed by Bonferroni’s post hoc test was used to analyze the data ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ns, non-significant ( p > 0.05).

Article Snippet: Rested immature dendritic cells were stimulated with 100 ng/mL of recombinant DKK1 (R & D Systems), 20 ng/mL of recombinant IL-10 (Thermo Fisher Scientific), or 10 ng/mL of recombinant TNF-α (Thermo Fisher Scientific).

Techniques: Infection, Saline, Isolation, Flow Cytometry

Journal: iScience

Article Title: Platelet DKK1 promotes tolerogenic dendritic cells and non-healing responses in cutaneous leishmaniasis

doi: 10.1016/j.isci.2026.115090

Figure Lengend Snippet: Increased CD38 + macrophages and CD8α + dendritic cells in MyD88 (PKO) - and DKK1 (PKO) -infected mice (A–D) Six-week-old female WT-BALB/c, MyD88 (PKO) and DKK1 (PKO) mice were challenged with infective metacyclic promastigote (2 ×10 6 parasites, n = 5 per group) of L . major via the footpad. One week and two weeks post-infection, cells from the infected foot of each mouse in BALB/c, MyD88 (PKO) , DKK1 (PKO) , and non-infected BALB/c mice ( n = 10/2 feet per mouse) were harvested and counted for macrophages (A and B) and dendritic cells (C and D) by flow cytometry. (E–H) Two weeks post-infection, the infected foot from each mouse in BALB/c, MyD88 (PKO) , DKK1 (PKO) ( n = 5 per group), and non-infected BALB/c mice ( n = 10/2 feet per mouse) were examined for CD206 + and CD38 + macrophages (E and F), as well as CD11b + and CD8α + dendritic cells (G and H) by flow cytometry. Representative flow cytometry dot plots showing the analysis of CD206 + , CD38 + macrophages and CD11b + , CD8α + dendritic cells and a dot plot of each sample in all the experimental groups performed on day 14 PI are presented in . Results are presented as mean ± SEM and are representative of two independent experiments. Data analysis was done using one-way ANOVA followed by Bonferroni’s post hoc test ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and “ns” indicates non-significant.

Article Snippet: Rested immature dendritic cells were stimulated with 100 ng/mL of recombinant DKK1 (R & D Systems), 20 ng/mL of recombinant IL-10 (Thermo Fisher Scientific), or 10 ng/mL of recombinant TNF-α (Thermo Fisher Scientific).

Techniques: Infection, Flow Cytometry

Journal: iScience

Article Title: Platelet DKK1 promotes tolerogenic dendritic cells and non-healing responses in cutaneous leishmaniasis

doi: 10.1016/j.isci.2026.115090

Figure Lengend Snippet: DKK1 promotes IL-10 induction in BALB/c-infected mice (A–F) Six-week-old female BALB/c, MyD88 (PKO) and DKK1 (PKO) mice were challenged with infective metacyclic promastigote (2 ×10 6 parasites, n = 5 per group) of L . major via the footpad. Non-infected BALB/c mice ( n = 5) were given 0.9% NaCl saline. Two or fifteen weeks post-infection, cells from draining lymph nodes were isolated. Lymph node cells were incubated with SLAG (50 μg/mL derived from WT parasites). Cell culture supernatant samples obtained were analyzed by ELISA for cytokine production, as shown in the column graphs (A–F). In all the experiments, BALB/c infected and non-infected mice served as positive and negative controls, respectively. Results are presented as mean ± SEM and are representative of two independent experiments. One-way ANOVA followed by Bonferroni’s post hoc test was used to analyze the data ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, “ns” indicates not significant ( p > 0.05).

Article Snippet: Rested immature dendritic cells were stimulated with 100 ng/mL of recombinant DKK1 (R & D Systems), 20 ng/mL of recombinant IL-10 (Thermo Fisher Scientific), or 10 ng/mL of recombinant TNF-α (Thermo Fisher Scientific).

Techniques: Infection, Saline, Isolation, Incubation, Derivative Assay, Cell Culture, Enzyme-linked Immunosorbent Assay

Journal: iScience

Article Title: Platelet DKK1 promotes tolerogenic dendritic cells and non-healing responses in cutaneous leishmaniasis

doi: 10.1016/j.isci.2026.115090

Figure Lengend Snippet: Impaired CD8 + IL-10 + IFN-γ − , CD4 + IL-10 + IFN-γ − , and CD4 + IL-10 + IFNg + T cells in MyD88 (PKO) - and DKK1 (PKO) -infected mice on day 14 PI (A–M) Six-week-old female BALB/c, MyD88 (PKO) , and DKK1 (PKO) mice were challenged with infective metacyclic promastigote (2 ×10 6 parasites, n = 5 per group) of L . major via the footpad. Non-infected BALB/c mice ( n = 5) were given 0.9% NaCl saline. Two weeks post-infection, the draining and non-draining lymph node cells were isolated. Lymph node cells were incubated with a cell stimulation cocktail for 5 h. After a further 3 h in BFA, cells were stained for intracellular IL-10 and IFN-γ. The stained lymph node cells from each mouse in BALB/c, MyD88 (PKO) - and DKK1 (PKO) -infected mice were determined for the percentage and frequency of CD4 + and CD8 + T cells (A, B, C, and B), percentage and MFI of CD8 + IFN-γ + or CD8 + IL-10 + T cells (E, F, G, and H), percentage and MFI of CD4 + IFN-γ + or CD4 + IL-10 + T cells (I, J, K, and L), and percentage of CD4 + IFN-γ + IL-10 + T cells (M) by flow cytometry. Representative flow cytometry dot plots showing the analyses CD4 + and CD8 + T cells, CD8 + IFN-γ + or CD8 + IL-10 + T cells, CD4 + IFN-γ + or CD4 + IL-10 + T cells, and CD4 + IFN-γ + IL-10 + T cells performed on day 14 PI is indicated ( A). Results are presented as mean ± SEM. and are representative of triplicate experiments. Data analysis was done using one-way ANOVA followed by Bonferroni’s post hoc test ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and “ns” indicates non-significant.

Article Snippet: Rested immature dendritic cells were stimulated with 100 ng/mL of recombinant DKK1 (R & D Systems), 20 ng/mL of recombinant IL-10 (Thermo Fisher Scientific), or 10 ng/mL of recombinant TNF-α (Thermo Fisher Scientific).

Techniques: Infection, Saline, Isolation, Incubation, Cell Stimulation, Staining, Flow Cytometry

Journal: iScience

Article Title: Platelet DKK1 promotes tolerogenic dendritic cells and non-healing responses in cutaneous leishmaniasis

doi: 10.1016/j.isci.2026.115090

Figure Lengend Snippet: Minimal expression and percentage of MHC II + , CD86 + , and CD80 + dendritic cells in the presence of recombinant DKK1 (A) Monocyte-derived dendritic cells were incubated in rDKK1(100 ng/mL), rIL-10 (20 ng/mL) and rTNF-α (10 ng/mL). Cells harvested at 24 and 48 h post-incubation were used to determine the expression and percentage of MHC II + , CD86 + , and CD80 + cells by flow cytometry. Representative flow cytometry dot plots generated 24 h post-incubation showed the analysis of MHC II + , CD86 + and CD80 + as indicated (A). (B–G) The percentage (B, D, and F) and MFI (C, E, and G) of MHC II + , CD86 + and CD80 + in the different experimental conditions are shown in the bar graphs. Results are presented as mean (±SEM) and are representative of triplicate experiments. One-way ANOVA followed by Bonferroni’s post hoc test was used to analyze the data (∗ p < 0.05, ∗∗ p < 0.01; ∗∗∗ p < 0.001; and “ns” indicates non-significant.

Article Snippet: Rested immature dendritic cells were stimulated with 100 ng/mL of recombinant DKK1 (R & D Systems), 20 ng/mL of recombinant IL-10 (Thermo Fisher Scientific), or 10 ng/mL of recombinant TNF-α (Thermo Fisher Scientific).

Techniques: Expressing, Recombinant, Derivative Assay, Incubation, Flow Cytometry, Generated

Journal: iScience

Article Title: Platelet DKK1 promotes tolerogenic dendritic cells and non-healing responses in cutaneous leishmaniasis

doi: 10.1016/j.isci.2026.115090

Figure Lengend Snippet: r-DKK1 blocked TNF-α-induced MHC II + and CD86 + expression of dendritic cells (A) Monocyte-derived dendritic cells were incubated in rDKK1(100 ng/mL), rTNF-α (10 ng/mL), and (rDKK1 + rTNF-α). Cells harvested at 48 h post-incubation were used to determine the percentage of MHC II + and CD86 + cells by flow cytometry. Representative flow cytometry dot plots generated 48 h post-incubation showed the analysis of MHC II + and CD86 + as indicated (A). (B and C) The percentage (B and C) of MHC II + and CD86 + in the different experimental conditions are shown in the bar graphs. Results are presented as mean (±SEM) and are representative of triplicate experiments. One-way ANOVA followed by Bonferroni’s post hoc test was used to analyze the data (∗ p < 0.05, ∗∗ p < 0.01; and “ns” indicates non-significant.

Article Snippet: Rested immature dendritic cells were stimulated with 100 ng/mL of recombinant DKK1 (R & D Systems), 20 ng/mL of recombinant IL-10 (Thermo Fisher Scientific), or 10 ng/mL of recombinant TNF-α (Thermo Fisher Scientific).

Techniques: Expressing, Derivative Assay, Incubation, Flow Cytometry, Generated

Journal: iScience

Article Title: Platelet DKK1 promotes tolerogenic dendritic cells and non-healing responses in cutaneous leishmaniasis

doi: 10.1016/j.isci.2026.115090

Figure Lengend Snippet: Elevated IL-10 and reduced IL-12 production in rDKK1-treated dendritic cells (A and B) Monocyte-derived dendritic cells were incubated in r-DKK1(100 ng/mL), r-IL-10 (20 ng/mL), r-TNF-α (10 ng/mL) or r-TNF-α + r-DKK1. Cell culture supernatants harvested 24 and 48 h post-incubation were used to determine IL-10 and IL-12 production by ELISA, as shown in the bar graphs (A and B). Results are presented as mean ± SEM and are representative of triplicate experiments. One-way ANOVA followed by Bonferroni’s post hoc test was used to analyze the data (∗ p < 0.05, ∗∗ p < 0.01). “ns” indicates not significant.

Article Snippet: Rested immature dendritic cells were stimulated with 100 ng/mL of recombinant DKK1 (R & D Systems), 20 ng/mL of recombinant IL-10 (Thermo Fisher Scientific), or 10 ng/mL of recombinant TNF-α (Thermo Fisher Scientific).

Techniques: Derivative Assay, Incubation, Cell Culture, Enzyme-linked Immunosorbent Assay

Journal: iScience

Article Title: Platelet DKK1 promotes tolerogenic dendritic cells and non-healing responses in cutaneous leishmaniasis

doi: 10.1016/j.isci.2026.115090

Figure Lengend Snippet: rDKK1-treated dendritic cells significantly increased the percentage of IL-10-producing CD4 T cells (A and B) Rested immature dendritic cells were stimulated with 100 ng/mL of recombinant DKK1 or 10 ng/mL of recombinant TNF-α for 24 h. The rDKK1 and r-TNF-α-treated dendritic cells were incubated for 4 h in a 24-well plate (2 × 10 5 per well) in 200 μL of medium in the presence of SLAG (100 ng/ml). SLAG-loaded rDKK1 and r-TNF-α-treated dendritic cells (2 ×10 5 ) were then washed and co-cultured with infected WT-BALB/c lymph node T cells (1 ×10 6 ) in complete RPMI 1640 medium in a 1:5 ratio for 72 h. The cells were surface-stained with appropriate antibodies before intracellular IL-10 staining with a cell stimulation cocktail and BD Golgi plug. The stained lymph node cells from the experimental condition were determined for the percentage of CD4 + IL-10 + T cells by flow cytometry. Representative flow cytometry dot plots showed the analysis of CD4 + IL-10 + T cells (A). The percentage of CD4 + IL-10 + T cells in the different experimental conditions is indicated (B). The non-treated and r-TNF-α-treated dendritic cells serve as controls. Results are presented as mean ± SEM and are representative of three replicate experiments. One-way ANOVA followed by Bonferroni’s post hoc test was used to analyze the data ∗∗∗ p < 0.001; ns indicates not significant ( p > 0.05).

Article Snippet: Rested immature dendritic cells were stimulated with 100 ng/mL of recombinant DKK1 (R & D Systems), 20 ng/mL of recombinant IL-10 (Thermo Fisher Scientific), or 10 ng/mL of recombinant TNF-α (Thermo Fisher Scientific).

Techniques: Recombinant, Incubation, Cell Culture, Infection, Staining, Cell Stimulation, Flow Cytometry

Journal: iScience

Article Title: Platelet DKK1 promotes tolerogenic dendritic cells and non-healing responses in cutaneous leishmaniasis

doi: 10.1016/j.isci.2026.115090

Figure Lengend Snippet: Lesion size and parasite burden decreased in MyD88 (PKO) - and DKK1 (PKO) -infected mice (A–C) Six-week-old female WT-BALB/c, MyD88 (PKO) and DKK1 (PKO) mice were challenged with infective metacyclic promastigote (2 ×10 6 parasites) of L . major via the footpad. The infected foot from each mouse in BALB/c, MyD88 (PKO) - and DKK1 (PKO) -infected mice ( n = 5 per group) were measured for lesion size weekly using a vernier caliper (A), and parasite burden (at day week 6 and 15PI) was determined by limiting dilution assay (B and C). Results are presented as mean ± SEM. For Figure (A), mice in each infected group were compared with the non-infected group and data analysis was done using one-way ANOVA followed by Bonferroni’s post hoc test ∗ p < 0.05; ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: Rested immature dendritic cells were stimulated with 100 ng/mL of recombinant DKK1 (R & D Systems), 20 ng/mL of recombinant IL-10 (Thermo Fisher Scientific), or 10 ng/mL of recombinant TNF-α (Thermo Fisher Scientific).

Techniques: Infection, Limiting Dilution 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