Journal: Medicina

Article Title: The Role of Serum Dickkopf1 and CKAP4 Levels in Diagnosing Colorectal Cancer and Measuring the Disease Severity: A Prospective Study

doi: 10.3390/medicina60060933

Figure Lengend Snippet: DKK1–CKAP4 serum levels according to stages in colorectal cancer patients.

Article Snippet: Serum obtained from whole blood samples collected at admission were analyzed via enzyme-linked immunosorbent assay (ELISA) using the Human CKAP4 ELISA Kit (BT LAB, Cat. No. E4664Hu, Shanghai, China) and the Human DKK1 ELISA Kit (BT LAB, Cat. No. E0630Hu, China) according to the manufacturer’s instructions.

Techniques:

Journal: Medicina

Article Title: The Role of Serum Dickkopf1 and CKAP4 Levels in Diagnosing Colorectal Cancer and Measuring the Disease Severity: A Prospective Study

doi: 10.3390/medicina60060933

Figure Lengend Snippet: Serum DKK1–CKAP4 levels according to grades in colorectal cancer patients.

Article Snippet: Serum obtained from whole blood samples collected at admission were analyzed via enzyme-linked immunosorbent assay (ELISA) using the Human CKAP4 ELISA Kit (BT LAB, Cat. No. E4664Hu, Shanghai, China) and the Human DKK1 ELISA Kit (BT LAB, Cat. No. E0630Hu, China) according to the manufacturer’s instructions.

Techniques:

Journal: Medicina

Article Title: The Role of Serum Dickkopf1 and CKAP4 Levels in Diagnosing Colorectal Cancer and Measuring the Disease Severity: A Prospective Study

doi: 10.3390/medicina60060933

Figure Lengend Snippet: CKAP4 and DKK1 levels of colorectal cancer and control group patients.

Article Snippet: Serum obtained from whole blood samples collected at admission were analyzed via enzyme-linked immunosorbent assay (ELISA) using the Human CKAP4 ELISA Kit (BT LAB, Cat. No. E4664Hu, Shanghai, China) and the Human DKK1 ELISA Kit (BT LAB, Cat. No. E0630Hu, China) according to the manufacturer’s instructions.

Techniques: Control

Journal: Medicina

Article Title: The Role of Serum Dickkopf1 and CKAP4 Levels in Diagnosing Colorectal Cancer and Measuring the Disease Severity: A Prospective Study

doi: 10.3390/medicina60060933

Figure Lengend Snippet: Correlation of DKK1-CKAP4 serum levels.

Article Snippet: Serum obtained from whole blood samples collected at admission were analyzed via enzyme-linked immunosorbent assay (ELISA) using the Human CKAP4 ELISA Kit (BT LAB, Cat. No. E4664Hu, Shanghai, China) and the Human DKK1 ELISA Kit (BT LAB, Cat. No. E0630Hu, China) according to the manufacturer’s instructions.

Techniques:

Journal: Medicina

Article Title: The Role of Serum Dickkopf1 and CKAP4 Levels in Diagnosing Colorectal Cancer and Measuring the Disease Severity: A Prospective Study

doi: 10.3390/medicina60060933

Figure Lengend Snippet: ROC curve results and sensitivity and specificity values of colorectal cancer patients and healthy controls.

Article Snippet: Serum obtained from whole blood samples collected at admission were analyzed via enzyme-linked immunosorbent assay (ELISA) using the Human CKAP4 ELISA Kit (BT LAB, Cat. No. E4664Hu, Shanghai, China) and the Human DKK1 ELISA Kit (BT LAB, Cat. No. E0630Hu, China) according to the manufacturer’s instructions.

Techniques:

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Enhancing motor functional recovery in spinal cord injury through pharmacological inhibition of Dickkopf-1 with BHQ880 antibody.

doi: 10.1016/j.biopha.2024.116792

Figure Lengend Snippet: Fig. 1. In vitro evaluation of the proper effect of the BHQ880 antibody over Dkk1 protein neutralization. (A) Representative images of B1a/7TGC cell reporter line activation when stimulated by the GSK-3 inhibitor CHIR99021 (positive control) or by increasing doses of recombinant mouse Wnt3a (mWnt3a), and data obtained from the evaluation of the percentage of GFP+ cells after 48 hours. *vs. Non-treated (NT); # vs. 0 ng/mL mWnt3a. (B) Representative images of B1a/7TGC cell reporter line inactivation when previously stimulated cells with 150 ng/mL of mWnt3a were turned off through the addition of increasing doses of recombinant rDkk1, and data obtained from the evaluation of the percentage of GFP+ cells after 48 hours. *vs. NT; # vs. 0 ng/mL rDkk1. (C) Representative images of the BHQ880 neutralizing effect over rDkk1 protein in the B1a/7TGC reporter cells stimulated with 150 ng/mL of mWnt3a, together with 250 ng/mL of rDkk1 and increasing doses of BHQ880 antibody, and data obtained from the evaluation of the percentage of GFP+ cells after 48 hours. *vs. NT; & vs. Wnt3a; # vs 0 ng/mL BHQ880. A one-way ANOVA followed by a Bonferroni post-hoc test was used to assess the existence of significant differences between groups. In all cases, scale bars = 50 μm and the data are presented as the mean + SEM. **p <0.01; ***p <0.001; #p <0.05; ##p <0.01; ###p <0.001; &&p <0.01.

Article Snippet: Finally, to validate the specificity of the Dkk1 primary antibody used in these experiments, we pre-adsorbed the antibody with its corresponding blocking peptide (Proteintech, AG15110) at a 20-fold weight/ weight excess following the protocol previously described [51], and we observed that pre-adsorption abolished Dkk1 immunostaining, thus confirming antibody specificity (Fig. S1).

Techniques: In Vitro, Neutralization, Activation Assay, Positive Control, Recombinant

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Enhancing motor functional recovery in spinal cord injury through pharmacological inhibition of Dickkopf-1 with BHQ880 antibody.

doi: 10.1016/j.biopha.2024.116792

Figure Lengend Snippet: Fig. 2. Protein expression of Dkk1 in the healthy and injured spinal cord. The figure shows representative images from the immunohistochemical evaluation of Dickkopf-1 (Dkk1) protein expression in the non-lesioned (NL) (n = 5) spinal cord (A) and after SCI at 24 hours post-injury (hpi) (n = 4), and 3 days post-injury (dpi), 7 dpi (n = 4), 14 dpi (n = 5), 28 dpi (n = 3), and 42 dpi (n = 5) at the lesion epicenter (B-G). Dashed line squares in A-G indicate the areas shown in the corresponding higher magnification images. Scale bars = 500 μm. Inserts scale bars = 50 μm. (H) Densitometric analysis of Dkk1 immunolabeling in the NL spinal cord and at the injury epicenter after SCI from 24 hpi to 42 dpi. A one-way ANOVA followed by a Bonferroni post-hoc test was used to assess the existence of significant differences between NL and the different times post-injury. In all cases, the data are presented as the mean + SEM. **p <0.01; ***p <0.001.

Article Snippet: Finally, to validate the specificity of the Dkk1 primary antibody used in these experiments, we pre-adsorbed the antibody with its corresponding blocking peptide (Proteintech, AG15110) at a 20-fold weight/ weight excess following the protocol previously described [51], and we observed that pre-adsorption abolished Dkk1 immunostaining, thus confirming antibody specificity (Fig. S1).

Techniques: Expressing, Immunohistochemical staining, Immunolabeling

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Enhancing motor functional recovery in spinal cord injury through pharmacological inhibition of Dickkopf-1 with BHQ880 antibody.

doi: 10.1016/j.biopha.2024.116792

Figure Lengend Snippet: Fig. 3. Analysis of the circulating levels of Dkk1 in Serum samples from non- lesioned (NL) and lesioned rats at 24 hours after damage (SCI). The figure shows data obtained from the quantitative analysis of serum circulating Dkk1 protein levels measured by ELISA in NL animals (n = 9) and injured animals 24 hours after SCI (n = 9). No significant differences were observed between groups. A two-tailed t test was used to determine the existence of significant differences between groups. The data represent the mean ± SEM.

Article Snippet: Finally, to validate the specificity of the Dkk1 primary antibody used in these experiments, we pre-adsorbed the antibody with its corresponding blocking peptide (Proteintech, AG15110) at a 20-fold weight/ weight excess following the protocol previously described [51], and we observed that pre-adsorption abolished Dkk1 immunostaining, thus confirming antibody specificity (Fig. S1).

Techniques: Enzyme-linked Immunosorbent Assay, Two Tailed Test

Journal: Frontiers in Oncology

Article Title: AXL Overexpression in Tumor-Derived Endothelial Cells Promotes Vessel Metastasis in Patients With Hepatocellular Carcinoma

doi: 10.3389/fonc.2021.650963

Figure Lengend Snippet: AXL/SOX2/DKK-1 axis in HUVECs promotes HCC metastasis. (A) DKK-1 and CCL14 secretion was significantly downregulated in CM from HUVEC-AXL-KD compared with that from HUVEC-AXL-NC, as detected with a human cytokine antibody array. (B) DKK-1 and CCL14 expression was markedly upregulated in the CM of HUVECs overexpressing AXL (CCL14: p < 0.001; DKK-1: p = 0.003) compared with the CM of HUVEC-AXL-NC, as detected by ELISA assay. (C) AXL siRNA downregulated DKK-1 and CCL14 secretion in the CM of HUVEC-AXL-NC and HUVEC-AXL-OE cells (CCL14: p < 0.001 and p < 0.001; DKK-1: p < 0.001 and p < 0.001). (D) DKK1 siRNA (MHCC-97L: p < 0.001 and p < 0.001; HCC-LM3: p <0.001 and p < 0.001), but not CCL14 siRNA (MHCC-97L: p = 0.126 and p = 0.711; HCC-LM3: p = 0.901 and p = 0.694) could attenuate the effect of the CM from HUVEC-AXL-NC and HUVEC-AXL-OE cells on the migration of HCC-LM3 cells and MHCC-97L cells. (E) SOX2 mRNA expression was significantly increased in HUVEC-AXL-OE cells and decreased in HUVEC-AXL-KD cells compared with HUVEC-AXL-NC cells (HUVEC-AXL-KD: p < 0.001, HUVEC-AXL-OE: p < 0.001). (F) AXL overexpression could significantly increase SOX2 and DKK-1 protein expression in HUVEC-AXL-OE cells compared with HUVEC-AXL-NC cells, and SOX2 siRNA inhibited SOX2 and DKK-1 protein expression in HUVEC-AXL-OE and HUVEC-AXL-NC cells.

Article Snippet: The protein concentrations of CCL14 and DKK-1 in the supernatants were also measured using an enzyme-linked immunosorbent assay (ELISA) kit (CCL14: EK1123 Boster, Wuhan, China; DKK-1: EK0867 Boster, Wuhan, China) according to the manufacturer’s instructions.

Techniques: Ab Array, Expressing, Enzyme-linked Immunosorbent Assay, Migration, Over Expression

Journal: Heliyon

Article Title: Potential actions of capsaicin for preventing vascular calcification of vascular smooth muscle cells in vitro and in vivo

doi: 10.1016/j.heliyon.2024.e28021

Figure Lengend Snippet: Cap inhibits Pi-induced calcification of VSMCs by inhibiting the Wnt/β-catenin signaling pathway by upregulation of TRPV1 receptor expression. A) RT-qPCR showing the mRNA levels of Wnt3a and β-catenin; B) Western blot illustrating the protein expression levels of Wnt3a and β-catenin; C) Alizarin Red staining assay; D) Calcium content in VSMCs assessed using a calcium colorimetric assay kit. E) RT-qPCR showing the expression of osteogenesis-specific and phenotypic markers in VSMC calcification. F) Western blot depicting the protein expression levels of osteogenesis-specific and phenotypic markers in VSMC calcification; G) Western blot showing the protein expression of β-catenin after treatment with DKK1. VSMCs cultured in the complete and pro-calcifying medium were defined as the control and Pi group, respectively; Pi + Cap: VSMCs cultured in the pro-calcifying medium with 20 μM Cap; Pi + Cap + CPZ: VSMCs cultured in the pro-calcifying medium with 20 μM Cap and 10 μM CPZ; Pi + Cap + DKK1: VSMCs cultured in the pro-calcifying medium with 20 μM Cap and 20 ng/mL DKK1; Data are mean ± SD (n = 3); #: p < 0.05, ##: p < 0.01, ###: p < 0.001 vs Control; *: p < 0.05, **: p < 0.01, ***: p < 0.001 vs Pi; ※: p < 0.05, ※※: p < 0.01, ※※※: p < 0.001 vs Pi + Cap; ns: p > 0.05 vs Control; p > 0.05 vs Pi; p > 0.05 vs Pi + Cap. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Antibodies against BMP-2 (PRP100126) and Runx2 (ABP53087) were purchased from Abbkine (Wuhan, Hubei, China); SM22α (AF9266) and Wnt3a (DF6113), Affinity Biosciences LTD; β-actin (81115-1-RR), proteintech (Hubei, China); β-catenin (ab32572), Abcom (USA); and DKK1 ( HY-P73305 ), MCE China (Beijing, China).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Staining, Calcium Colorimetric Assay, Cell Culture, Control

Journal: Natural Products and Bioprospecting

Article Title: Scutellarin ameliorates diabetic nephropathy via TGF-β1 signaling pathway

doi: 10.1007/s13659-024-00446-y

Figure Lengend Snippet: Scutellarin Restored Podocyte Injury of the DN Mice. a Representative images of immunohistochemistry for NPHS1 and NPHS2 of the mice treated with vehicle, scutellarin or empagliflozin (× 200; scale bar = 50 µm). b Representative images of Western-blotting for NPHS1, NPHS2. c Quantitative plot of the expression of NPHS1 of the mice. d Quantitatification of NPHS1 expression of the mice. e Representative images of Western-blotting for β-catenin, Axin2, snail and DKK1 of the mice. f – i Quantifications of the protein levels for β-catenin, Axin2, snail and DKK1 from E. All data are presented as the mean ± S.D.; n = 4–6 for each group, “n” stands for the number of animals; p vs. the model group (STZ)

Article Snippet: Scutellarin (Yunnan Phytopharmaceutical Co., LTD., China); Empagliflozin (Cat. C14295412, Macklin Biochemical, China), Streptozotocin (Cat. S8050, Solarbio, China); goat anti-rabbit immunoglobulin G (IgG) horseradish peroxidase (HRP)-linked antibody (Cat. AS014, Abclonal, China); anti-mouse IgG HRP-linked antibody (Cat. 7076S, Cell Signaling Technology, USA); Methenamine Silver Plating Stain Kit (Cat. G1790, Solarbio); Glycogen Periodic Acid Schiff (PAS) Stain Kit (Cat. G1281, Solarbio); Masson’s Trichrome Stain Kit (Cat. G1340, Solarbio); Mouse MAU enzyme-linked immunosorbent assay (ELISA) Kit (Cat. JL20493, JONIN, China); Anti-DKK1 antibody (Cat. A00632, Boster Biological Technology, China); Anti-SNAIl antibody (Cat. BP0449, Boster Biological Technology); Anti-α-SMA antibody (Cat. BM0002, Boster Biological Technology); Anti-TGFB1 antibody (Cat. BA0290, Boster Biological Technology); Anti-NPHS1 antibody (Cat. BA1669, Boster Biological Technology); Anti-NPHS2 antibody (Cat. BA1688, Boster Biological Technology); Anti-SMAD2/3 antibody (Cat. BA1395, Boster Biological Technology); Anti-Phospho-SMAD2(s250) antibody (Cat. BM4693, Boster Biological Technology); Anti-FN1 antibody (Cat. BA1772, Boster Biological Technology); AXIN2 antibody (Cat. Ab32197, Abcam, USA); Anti-Phospho-SMAD3(ser425) antibody (Cat. AF3362, Affinity Biosciences, China); Anti-COL3A1 antibody (Cat. M00788, Boster Biological Technology); Anti-extracellular signal-regulated kinase (ERK)1/2 antibody (Cat. ET1601-29, HUABIO, China); Anti-ERK1(PT202/PY204) + ERK2(PT185/PY187) antibody (Cat. ET1610-13, HUABIO); Anti-P38 antibody (Cat. ET1602-26, HUABIO); Anti-Phospho-P38(Thr180 + Tyr182) antibody (Cat. ER1903-01, HUABIO); Anti-β-Actin antibody (Cat. EM21002, HUABIO); Anti-SMAD4 antibody (Cat. A5657, Abclonal); Anti-β-catenin antibody (Cat. 610154, BD Biosciences, China); Creatinine (Cr) Assay kit (Cat. C011-2-1, Nanjing Jiancheng Bioengineering Institute, China); Glucose Assay Kit (Cat. F006-1-1, Nanjing Jiancheng Bioengineering Institute); microalbunminuria (MAU) ELISA kit (Cat. JL20493, JONIN).

Techniques: Immunohistochemistry, Western Blot, Expressing