Journal: Nature communications

Article Title: Stroma-derived Dickkopf-1 contributes to the suppression of NK cell cytotoxicity in breast cancer.

doi: 10.1038/s41467-025-56420-w

Figure Lengend Snippet: Fig. 8 | DKK1 levels correlate with reduced therapeutic responses and cytotoxic NK cells. A Schematic representation of NK cell isolation from healthy donor human PBMCs and incubation with target cells in the presence of PBS or rhDKK1 for 4 h. B–D Analysis of percent specific killing measured by 7-AAD+ MDA-MB-231 (B, n = 7 NK donors), T47D (C, n = 3 NK donors), and K562 (D, n = 3 NK donors) target tumor cells after 4 h incubation with human NK cells in the presence of rhDKK1 or PBS as a control. E FACS analysis of NK cell activating receptors on human NK cells from different donors (n = 11) stimulated with rhDKK1 for 24 h or PBS. F Schematic representation of blood sample collection of advanced breast

Article Snippet: Serum or plasma levels of DKK1 frommice or patients were quantified using ELISA kits specific for DKK1 (Mouse Dkk1 Quantikine ELISA kit or Human Dkk1 Quantikine ELISA kit, R&D systems) as per the manufacturer’s protocols.

Techniques: Cell Isolation, Incubation, Control

Journal: Journal of the American Society of Nephrology : JASN

Article Title: CKD-Induced Wingless/Integration1 Inhibitors and Phosphorus Cause the CKD–Mineral and Bone Disorder

doi: 10.1681/ASN.2013080818

Figure Lengend Snippet: Experimental design. (A) Schematic drawing of the experimental design defining the various animal groups. Wild-type C57B6J mice (WT) were used to establish the normal ranges of the parameters being studied. CKD-2 was induced in ldlr−/− mice fed high-fat diets (d) at 10 weeks of life by electrocautery (EC) injury of one kidney at 12 weeks and contralateral nephrectomy (Nx) at 14 weeks. The mice received vehicle injections from 14 weeks to euthanasia at 22 weeks. The Dkk1 mAb-treated mice (CKD-2/Dkk1 mAb) were ldlr−/− high fat-fed mice undergoing the same protocol, except for receiving the Dkk1 mAb (30 mg/kg intraperitoneally twice weekly) instead of vehicle. The control mice for the effects of early CKD were ldlr−/− mice in the C57B6J background fed a high-fat diet undergoing sham operations (SOs). (B) GFRs by inulin clearance in Sham, CKD-2, and CKD-2/Dkk1 mAb-treated mice. Inulin clearances and BUN levels (Table 1) were used to establish that the mild ablative CKD was equivalent in GFR reduction to human stage 2 CKD. Inulin clearances were at 20 weeks of age. The reduction in GFR was equivalent to the reduction in eGFR considered as stage 2 CKD clinically. Dkk1 mAb treatment did not affect GFR.

Article Snippet: Serum Dkk1 levels were analyzed using a commercial mouse Dkk1 ELISA kit (catalog number DY1765; R&D Systems, Inc., Minneapolis, MN).

Techniques: Control

Journal: Journal of the American Society of Nephrology : JASN

Article Title: CKD-Induced Wingless/Integration1 Inhibitors and Phosphorus Cause the CKD–Mineral and Bone Disorder

doi: 10.1681/ASN.2013080818

Figure Lengend Snippet: Kidney homogenate and plasma Dkk1 levels. (A) Dkk1 levels in kidney homogenates were increased at 15 weeks in CKD-2 mice. By 22 weeks, kidney Dkk1 levels in the CKD-2 and CKD-2+Dkk1 mAb mice were similar to sham mice. The increased Dkk1 expression in sham-operated mice compared with wild-type mice is related to diabetes. IB, immunoblot. (B) Plasma Dkk1 levels were massively increased in the AKI phase at 15 weeks and remained 4-fold elevated at 22 weeks. Compared with wild-type mice, the sham mice did not have increased Dkk1 in the plasma, despite the increased protein levels in renal homogenates. The Dkk1 mAb affected the ELISA assay for Dkk1, preventing measurement of plasma levels in the Dkk1 mAb-treated group.

Article Snippet: Serum Dkk1 levels were analyzed using a commercial mouse Dkk1 ELISA kit (catalog number DY1765; R&D Systems, Inc., Minneapolis, MN).

Techniques: Clinical Proteomics, Expressing, Western Blot, Enzyme-linked Immunosorbent Assay

Journal: Journal of the American Society of Nephrology : JASN

Article Title: CKD-Induced Wingless/Integration1 Inhibitors and Phosphorus Cause the CKD–Mineral and Bone Disorder

doi: 10.1681/ASN.2013080818

Figure Lengend Snippet: Effect of the Dkk1 neutralizing antibody on skeletal histomorphometry and μCT imaging of femoral metaphyseal trabeculae and midshaft femur cortices. (A) Histomorphometry: Dkk1 antibody treatment increased trabecular bone volume (BV/TV), osteoblast number (ObN/BL), and osteoclast number (OcN/BL) compared with CKD-2 vehicle-treated mice (OV/BV). (B) Dkk1 antibody treatment increased bone formation rates (BFRs). (C) μCT of femoral metaphyseal trabecular bone revealed increased bone volume (BV/TV), trabecular number (Tb.N), and decreased trabecular separation (Tb.Sp) in the Dkk1 mAb-treated mice without changing trabecular thickness (Tb.Th). There were no significant differences between sham-operated and CKD-2 mice. wild-type (WT), n=11; sham, n=9; CKD-2, n=11; Dkk1 Ab, n=12. *P<0.05; **P<0.005. (D) Three-dimensional reconstruction of the distal femoral cortical bone showing loss of bone volume in CKD-2 mice prevented by treatment with the Dkk1 mAb. Data are shown as mean±SEM.

Article Snippet: Serum Dkk1 levels were analyzed using a commercial mouse Dkk1 ELISA kit (catalog number DY1765; R&D Systems, Inc., Minneapolis, MN).

Techniques: Imaging

Journal: Journal of the American Society of Nephrology : JASN

Article Title: CKD-Induced Wingless/Integration1 Inhibitors and Phosphorus Cause the CKD–Mineral and Bone Disorder

doi: 10.1681/ASN.2013080818

Figure Lengend Snippet: Aortic calcification and osteoblastic transition in the various groups of mice. (A) Aortic calcium was increased in the CKD-2 mice compared with sham and wild-type mice. The increase in aortic Ca was prevented by treatment with the Dkk1 antibody. (B) Relative mRNA levels of smooth muscle 22α (SM22α), ALP, Runx2, and FGF23. In the two bars on the left of the graphs, the effect of CKD-2 on the message levels is compared with levels in aortas of sham-operated mice, which were set to equal to one. Then, in the bars on the right, the aortic message levels in mice treated with the Dkk1 antibody are compared with levels in vehicle-treated CKD-2 mice, which were set to equal to one. (C) Immunoblots of aortic α-smooth muscle actin, Runx2, and FGF23; α-tubulin levels served as loading controls. IB, immunoblot. *P<0.05.

Article Snippet: Serum Dkk1 levels were analyzed using a commercial mouse Dkk1 ELISA kit (catalog number DY1765; R&D Systems, Inc., Minneapolis, MN).

Techniques: Western Blot

Journal: Journal of the American Society of Nephrology : JASN

Article Title: CKD-Induced Wingless/Integration1 Inhibitors and Phosphorus Cause the CKD–Mineral and Bone Disorder

doi: 10.1681/ASN.2013080818

Figure Lengend Snippet: Effect of Dkk1 neutralizing antibody on aortic and circulating klotho levels. (A) Aortic mRNA and immunoblots for αklotho. (B) Aortic immunohisotochemistry for αklotho. (C) Plasma cklotho levels in the various groups of mice at euthanasia. IB, immunoblot; WT, wild-type.

Article Snippet: Serum Dkk1 levels were analyzed using a commercial mouse Dkk1 ELISA kit (catalog number DY1765; R&D Systems, Inc., Minneapolis, MN).

Techniques: Western Blot, Clinical Proteomics

Journal: Journal of the American Society of Nephrology : JASN

Article Title: CKD-Induced Wingless/Integration1 Inhibitors and Phosphorus Cause the CKD–Mineral and Bone Disorder

doi: 10.1681/ASN.2013080818

Figure Lengend Snippet: Effect of Dkk1 neutralizing antibody on osteocyte-secreted proteins FGF23 and sclerostin (Scl). (A) Plasma FGF23 levels in the various groups of mice. (B) The change in Scl levels between weeks 15 and 22 of life in plasma of CKD-2 vehicle-treated mice and CKD-2 mice treated with Dkk1 mAb.

Article Snippet: Serum Dkk1 levels were analyzed using a commercial mouse Dkk1 ELISA kit (catalog number DY1765; R&D Systems, Inc., Minneapolis, MN).

Techniques: Clinical Proteomics

Journal: Journal of the American Society of Nephrology : JASN

Article Title: CKD-Induced Wingless/Integration1 Inhibitors and Phosphorus Cause the CKD–Mineral and Bone Disorder

doi: 10.1681/ASN.2013080818

Figure Lengend Snippet: Combination Dkk1 antibody and phosphate binder therapy. (A) Schematic drawing of the modified experimental design defining the various animal groups and a switch from a prevention to a treatment strategy. Sham, ldlr−/− mice on a high-fat diet undergoing SOs are the control mice for the effects of CKD; CKD-2–28, ldlr−/− high fat-fed mice undergoing mild renal injury (EC) and contralateral NX with vehicle treatment beginning at 22 weeks and euthanasia at 28 weeks; CKD-2–28-Dkk1 mAb, same as CKD-2–28 but with Dkk1 mAb (30 mg/kg intraperitoneally twice weekly) beginning at 22 weeks; CKD-2–28-LaCO3, same as CKD-2–28 but 3% LaCO3 added to food beginning at 22 weeks; CKD-2–28-Dkk1 mAb+LaCO3, same as CKD-2–28 but both the Dkk1 mAb and LaCO3 added at 22 weeks. (B) Inulin clearances in the mice used in the modified experimental design. (C) Effect of LaCO3 on plasma FGF23 levels. (D) Urine phosphate/creatinine ratios at euthanasia in the various groups of mice. Urinary phosphorus/creatinine was decreased by LaCO3 treatment. EC, electrocautery; SOs, sham operations; WT, wild-type. *P<0.05 compared to CKD-2 (22 weeks).

Article Snippet: Serum Dkk1 levels were analyzed using a commercial mouse Dkk1 ELISA kit (catalog number DY1765; R&D Systems, Inc., Minneapolis, MN).

Techniques: Modification, Control, Clinical Proteomics

Journal: Journal of the American Society of Nephrology : JASN

Article Title: CKD-Induced Wingless/Integration1 Inhibitors and Phosphorus Cause the CKD–Mineral and Bone Disorder

doi: 10.1681/ASN.2013080818

Figure Lengend Snippet: Serum chemistries in the modified study design

Article Snippet: Serum Dkk1 levels were analyzed using a commercial mouse Dkk1 ELISA kit (catalog number DY1765; R&D Systems, Inc., Minneapolis, MN).

Techniques: Modification

Journal: Journal of the American Society of Nephrology : JASN

Article Title: CKD-Induced Wingless/Integration1 Inhibitors and Phosphorus Cause the CKD–Mineral and Bone Disorder

doi: 10.1681/ASN.2013080818

Figure Lengend Snippet: Plasma FGF23, sclerostin, and Dkk1 levels in normophosphatemic patients with CKD-3a. Plasma levels were determined at baseline before treatment in 38 patients enrolled in a pilot and feasibility study of phosphate binding in normophosphatemic patients with early CKD. The levels in the CKD-3a cohort were compared with those levels in 700 patients with normal kidney function in the Diabetes Heart Study (DHS) determined simultaneously in the same assays. *P<0.01 compared with the DHS study cohort.

Article Snippet: Serum Dkk1 levels were analyzed using a commercial mouse Dkk1 ELISA kit (catalog number DY1765; R&D Systems, Inc., Minneapolis, MN).

Techniques: Clinical Proteomics, Binding Assay