Journal: International journal of molecular sciences

Article Title: LRP4 and Agrin Are Modulated by Cartilage Degeneration and Involved in β-Catenin Signaling in Human Articular Chondrocytes.

doi: 10.3390/ijms26031007

Figure Lengend Snippet: Figure 6. Western blot analysis of the effect of rhAgrin treatment on human articular chondrocytes. (A) At the whole protein level, the expression of β-catenin, RUNX2, and SOX9 were increased after rhAgrin treatment. (B) Analysis of nuclear extraction protein also showed that the expression of both β-catenin and SOX9 were increased after rhAgrin treatment. In contrast, analysis of nuclear extraction protein showed no significant difference in the expression of NF-κB p65. * p < 0.05, ** p < 0.01, ns: not significant.

Article Snippet: The membranes were incubated with Odyssey® Blocking Buffer (LI-COR Biosciences, Lincoln, NE, USA) for 1 h at room temperature and then incubated overnight at 4 ◦C with anti-SOX9 antibody (1:5000, ab185230, Abcam, Cambridge, UK), anti-β-catenin antibody (1:1000, #37447, Cell Signaling Technology), anti-RUNX2 antibody (1:1000, #12556, Cell Signaling Technology), or anti-NF-kB p65 (1:1000, #8242, Cell Signaling Technology) as the primary antibody.

Techniques: Western Blot, Expressing, Extraction

Journal: International journal of molecular sciences

Article Title: LRP4 and Agrin Are Modulated by Cartilage Degeneration and Involved in β-Catenin Signaling in Human Articular Chondrocytes.

doi: 10.3390/ijms26031007

Figure Lengend Snippet: Figure 5. Immunocytochemical evaluation of the effect of rhAgrin treatment on human articular chondrocytes. (A) Double immunocytochemical staining of β-catenin and SOX9 nuclear translocation. (B) Translocation to nuclei of both β-catenin and SOX9 was increased after rhAgrin treatment. * p < 0.001. Scale bar = 100 µm.

Article Snippet: The membranes were incubated with Odyssey® Blocking Buffer (LI-COR Biosciences, Lincoln, NE, USA) for 1 h at room temperature and then incubated overnight at 4 ◦C with anti-SOX9 antibody (1:5000, ab185230, Abcam, Cambridge, UK), anti-β-catenin antibody (1:1000, #37447, Cell Signaling Technology), anti-RUNX2 antibody (1:1000, #12556, Cell Signaling Technology), or anti-NF-kB p65 (1:1000, #8242, Cell Signaling Technology) as the primary antibody.

Techniques: Staining, Translocation Assay

Journal: International journal of molecular sciences

Article Title: LRP4 and Agrin Are Modulated by Cartilage Degeneration and Involved in β-Catenin Signaling in Human Articular Chondrocytes.

doi: 10.3390/ijms26031007

Figure Lengend Snippet: Figure 7. Combined effects of Agrin knockdown and intense CTS on β-catenin expression analyzed by Western blot. Expression of β-catenin was increased 24 h after application of intense CTS, but the siAGRN transfection + intense CTS group showed a trend towards reduced nuclear expression of β-catenin. * p < 0.001, ** p < 0.0001.

Article Snippet: The membranes were incubated with Odyssey® Blocking Buffer (LI-COR Biosciences, Lincoln, NE, USA) for 1 h at room temperature and then incubated overnight at 4 ◦C with anti-SOX9 antibody (1:5000, ab185230, Abcam, Cambridge, UK), anti-β-catenin antibody (1:1000, #37447, Cell Signaling Technology), anti-RUNX2 antibody (1:1000, #12556, Cell Signaling Technology), or anti-NF-kB p65 (1:1000, #8242, Cell Signaling Technology) as the primary antibody.

Techniques: Knockdown, Expressing, Western Blot, Transfection

Journal: International journal of molecular sciences

Article Title: LRP4 and Agrin Are Modulated by Cartilage Degeneration and Involved in β-Catenin Signaling in Human Articular Chondrocytes.

doi: 10.3390/ijms26031007

Figure Lengend Snippet: Figure 10. Modulation of LRP4 and Agrin under the current experimental conditions focusing on β-catenin signaling in human articular chondrocytes. Mild mechanical stress upregulated (↑) LRP4 expression in the superficial layer. Intense mechanical stress increased (↑) Agrin expression, leading to situations similar to LRP4 downregulation or LRP4 knockdown (↓), and then upregulated (↑) β-catenin translocation via LRP5/6. The LRP4 knockdown experiments showed that LRP4 negatively regulated LRP5/6 expression. FZD—Frizzled; SOST—Sclerostin; DKK1—Dickkopf-1.

Article Snippet: The membranes were incubated with Odyssey® Blocking Buffer (LI-COR Biosciences, Lincoln, NE, USA) for 1 h at room temperature and then incubated overnight at 4 ◦C with anti-SOX9 antibody (1:5000, ab185230, Abcam, Cambridge, UK), anti-β-catenin antibody (1:1000, #37447, Cell Signaling Technology), anti-RUNX2 antibody (1:1000, #12556, Cell Signaling Technology), or anti-NF-kB p65 (1:1000, #8242, Cell Signaling Technology) as the primary antibody.

Techniques: Expressing, Knockdown, Translocation Assay