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: 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

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