Journal: The Journal of Biological Chemistry

Article Title: Systematic Mapping of WNT-FZD Protein Interactions Reveals Functional Selectivity by Distinct WNT-FZD Pairs *

doi: 10.1074/jbc.M114.612648

Figure Lengend Snippet: WNT-3A, -4, and -5A induce dose-dependent phosphorylation and shift of DVL3 (PS-DVL3) in mouse 32D/FZD cells. A, the bar graphs show phosphorylation of DVL3 (PS-DVL3) in lysates from 32D/FZD2/4/5 cells stimulated for 2 h with PBS and 3, 10, 30, 100, 300, and 1000 ng/ml WNTs. Densitometry data ratios (i.e. the quotient of the values from the upper (PS-DVL) and lower DVL bands) were normalized to the unstimulated control values. Basal PS-DVL2 was not detectable (ND) in the parental 32D cells. B, representative immunoblots for detection of PS-DVL3. ▶, DVL3; ▷, PS-DVL3. β-Actin was used as a loading control in all the experiments. Error bars indicate standard S.E. (n ≥ 3). *, p < 0.05; **, p < 0.01; ***, p < 0.001.

Article Snippet: After blocking with 3% milk in Tris-buffered saline with 0.05% Tween 20, membranes were incubated in blocking buffer overnight at 4 °C with the following primary antibodies: mouse anti-β-catenin (1:1000; BD Biosciences 610153), mouse anti-DVL3 (1:500; Santa Cruz Biotechnology sc-8027), rabbit anti-DVL2 (1:1000; Cell Signaling Technology 3216), rabbit anti-phospho-LRP6 (1:1000; Cell Signaling Technology 2568), mouse anti-HA tag (1:1000; Abcam 9110), and anti-mouse β-actin (1:30,000; Sigma A5441).

Techniques: Western Blot

Journal: The Journal of Biological Chemistry

Article Title: Systematic Mapping of WNT-FZD Protein Interactions Reveals Functional Selectivity by Distinct WNT-FZD Pairs *

doi: 10.1074/jbc.M114.612648

Figure Lengend Snippet: Overview of the data presented in this study, including WNT-FZD CRD binding data and the functional response upon WNT-FZD binding ND, no activity or binding detectable with the methodology used.

Article Snippet: After blocking with 3% milk in Tris-buffered saline with 0.05% Tween 20, membranes were incubated in blocking buffer overnight at 4 °C with the following primary antibodies: mouse anti-β-catenin (1:1000; BD Biosciences 610153), mouse anti-DVL3 (1:500; Santa Cruz Biotechnology sc-8027), rabbit anti-DVL2 (1:1000; Cell Signaling Technology 3216), rabbit anti-phospho-LRP6 (1:1000; Cell Signaling Technology 2568), mouse anti-HA tag (1:1000; Abcam 9110), and anti-mouse β-actin (1:30,000; Sigma A5441).

Techniques: Binding Assay, Functional Assay, Activity Assay

Journal: Biological procedures online

Article Title: Circadian gene CSNK1D promoted the progression of hepatocellular carcinoma by activating Wnt/β-catenin pathway via stabilizing Dishevelled Segment Polarity Protein 3.

doi: 10.1186/s12575-022-00183-x

Figure Lengend Snippet: Fig. 7 Overexpression of CSNK1D promote the tumor growth and metastasis in vivo. A The effect of CSNK1D on tumor growth were evaluated by xenograft tumor in nude mice. B, C Volume and weight of xenograft tumors. D The effect of CSNK1D on tumor metastasis was observed in lung metastasis model. E H&E staining of the lung metastatic nodules. F The representative immunohistochemical staining (Ki67, β-catenin, N-cadherin, Vimentin, E-cadherin, DVL3) of the xenograft tumors

Article Snippet: The primary antibodies were diluted as follows: β-catenin (1:50, #8480, CST, USA), CSNK1D (1:50, #sc-55553, SantaCruz, USA), DVL3 (1:100, #13,444–1-AP, Proteintech, China).

Techniques: Over Expression, In Vivo, Staining, Immunohistochemical staining