rat anti wnt3a antibody  (R&D Systems)

Journal: Scientific Reports

Article Title: Glucose induced activation of canonical Wnt signaling pathway in hepatocellular carcinoma is regulated by DKK4

doi: 10.1038/srep27558

Figure Lengend Snippet: ( A ) ELISA measurements of Wnt3a and DKK4 secretory proteins in culture media collected after 16 hr from HepG2 cells in NG, HG and HG + CytoB. ( B ) HepG2 and SK-HEP-1 cells were cultured in NG, HG and HG + CytoB for 16 hr. Whole cell lysates were subjected to western blotting and levels of Wnt3a and DKK4 proteins were detected. ( C ) HepG2 cells were cultured in NG, HG and HG + CytoB for 16 hr. Total RNA was isolated and cDNA was prepared to determine relative mRNA fold expression of DKK4 by quantitative real-time RT-PCR. ( D ) HepG2 cells were cultured in HG for 16 hr and then allowed to grow in medium without glucose for indicated time course. Whole cell lysates were prepared for detection of DKK4 protein by western blotting. All the bar graphs represent the mean ± SD of an experiment done in triplicate (*P < 0.05, **P < 0.001, ***P < 0.0001). Cropped blots are used in the main figure and full length blots are included in .

Article Snippet: Antibodies against DKK4 (sc-25519), GLUT-1 (sc-7903), GLUT-2 (sc-9117), Cyclin D1(sc-717), CDK4 (sc-260), CDK6 (sc-7181), c-Myc (sc-764), Wnt3a (sc-28824), pJNK (sc-6254), JNK (sc-474), pMEK (sc-7995), MEK (sc-13069), Raf (sc-227), pERK (sc-7383), ERK (sc-154), pAkt (sc-7985), AKT (sc-8312), GSK3β (sc-9166), β-tubulin (sc-9104) and Histone H1 (sc-10806), HRP-conjugated (sc-2004), (sc-2031), (sc-2033), FITC conjugated (sc-3839) and Rhodamine conjugated (sc-358922) antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz Biotechnology, CA, USA).

Techniques: Enzyme-linked Immunosorbent Assay, Cell Culture, Western Blot, Isolation, Expressing, Quantitative RT-PCR

Journal: Scientific Reports

Article Title: Glucose induced activation of canonical Wnt signaling pathway in hepatocellular carcinoma is regulated by DKK4

doi: 10.1038/srep27558

Figure Lengend Snippet: ( A ) Normoglycemic glucose promotes sustained expression of DKK4 protein. DKK4 antagonizes activation of canonical Wnt signaling by facilitating degradation of β–catenin in cytosol and thus reducing its transcriptional activation thereby causing decrease c-Myc level. Increased DKK4 expression affects progression of cells at S-phase of cell cycle and therefore limits proliferation of HCC cells. ( B ) High glucose diminishes DKK4 expression allowing activation of canonical Wnt signaling because of inactivation of β–catenin degradation complex, by Wnt3a proteins. Increase in β–catenin level enhances its transcriptional activity and promotes c-Myc expression which causes uncontrolled proliferation of HCC cells.

Article Snippet: Antibodies against DKK4 (sc-25519), GLUT-1 (sc-7903), GLUT-2 (sc-9117), Cyclin D1(sc-717), CDK4 (sc-260), CDK6 (sc-7181), c-Myc (sc-764), Wnt3a (sc-28824), pJNK (sc-6254), JNK (sc-474), pMEK (sc-7995), MEK (sc-13069), Raf (sc-227), pERK (sc-7383), ERK (sc-154), pAkt (sc-7985), AKT (sc-8312), GSK3β (sc-9166), β-tubulin (sc-9104) and Histone H1 (sc-10806), HRP-conjugated (sc-2004), (sc-2031), (sc-2033), FITC conjugated (sc-3839) and Rhodamine conjugated (sc-358922) antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz Biotechnology, CA, USA).

Techniques: Expressing, Activation Assay, Activity Assay