Journal: Annals of Gastroenterological Surgery

Article Title: Cytoplasmic localization of connexin 26 suppresses transition of β‐catenin into the nucleus in intestinal‐ and mix‐type gastric cancer

doi: 10.1002/ags3.12552

Figure Lengend Snippet: Representative images of immunohistochemical staining of Cx26, β‐catenin, and Wnt3a in gastric cancer. 200‐fold magnification. A: Cx26 expression in noncancerous regions; B: low cytoplasmic Cx26 expression in cancerous regions; C: high cytoplasmic Cx26 expression in cancerous regions; D: low nuclear β‐catenin expression in cancerous regions; E: high nuclear β‐catenin expression in cancerous regions; F: low nuclear Wnt3a expression in cancerous regions; G: high Wnt3a expression in cancerous regions

Article Snippet: Anti‐Non‐phospho β‐catenin monoclonal antibody (1:600 dilution), and the Wnt3a antibody was Wnt3a (Bioss Antibodies, MA, USA), Wnt3a Polyclonal Antibody (1:200 dilution).

Techniques: Immunohistochemical staining, Staining, Expressing

Journal: Annals of Gastroenterological Surgery

Article Title: Cytoplasmic localization of connexin 26 suppresses transition of β‐catenin into the nucleus in intestinal‐ and mix‐type gastric cancer

doi: 10.1002/ags3.12552

Figure Lengend Snippet: Evaluation of cytoplasmic Cx26 expression and clinicopathological factors in both intestinal‐ and mix‐type and diffuse type gastric cancer

Article Snippet: Anti‐Non‐phospho β‐catenin monoclonal antibody (1:600 dilution), and the Wnt3a antibody was Wnt3a (Bioss Antibodies, MA, USA), Wnt3a Polyclonal Antibody (1:200 dilution).

Techniques: Expressing

Journal: Annals of Gastroenterological Surgery

Article Title: Cytoplasmic localization of connexin 26 suppresses transition of β‐catenin into the nucleus in intestinal‐ and mix‐type gastric cancer

doi: 10.1002/ags3.12552

Figure Lengend Snippet: Kaplan–Meier analysis of 5‐y overall survival. (A) Prognosis based on cytoplasmic Cx26 expression in intestinal and mix‐type gastric cancer. (B) Prognosis based on cytoplasmic Cx26 expression in diffuse type gastric cancer. (C) Prognosis based on cytoplasmic Cx26 expression when Wnt3a expression is low. (D) Prognosis based on cytoplasmic Cx26 expression when Wnt3a expression is high

Article Snippet: Anti‐Non‐phospho β‐catenin monoclonal antibody (1:600 dilution), and the Wnt3a antibody was Wnt3a (Bioss Antibodies, MA, USA), Wnt3a Polyclonal Antibody (1:200 dilution).

Techniques: Expressing

Journal: Stem cells (Dayton, Ohio)

Article Title: Activation of Wnt signaling in hematopoietic regeneration.

doi: 10.1634/stemcells.2007-0768

Figure Lengend Snippet: Figure 2. Wnt10b is temporally regulated in the bone marrow microenvironment after injury. (A): Bone sections of control or Cy/G-injured mice were stained for the expression of Wnt3a, Wnt5a, or Wnt10b (red) and 4,6-diamidino-2-phenylindole (blue) (n 2 for Wnt3a and Wnt5a and n 5 for Wnt10b). (B): Kinetics of Wnt10b expression levels were monitored for 1 week following injury by analyzing bone sections at days 3 (n 3), 5 (n 2), and 7 (n 3) after Cy/G delivery. All images were acquired with a 40 objective.

Article Snippet: Primary antibodies were as follows: Wnt3a-specific antisera from immunized rabbits [28] (generous gift of Roel Nusse), anti-Wnt5a (Santa Cruz Biotechnology Inc., Santa Cruz, CA, http:// www.scbt.com), anti-Wnt10b (Santa Cruz), and/or biotin-conjugated mouse panendothelial cell antigen (MECA)-32 (BD Pharmingen) or the appropriate isotype controls; normal rabbit serum (Jackson Immunoresearch Laboratories), normal goat IgG (Santa Cruz), or biotin-conjugated rat IgG2a,k (BD Pharmingen).

Techniques: Control, Staining, Expressing

Journal: Journal of Functional Foods

Article Title: Thelephoric acid, p-terphenyl, induces bone-forming activities in pre-osteoblasts

doi: 10.1016/j.jff.2022.105036

Figure Lengend Snippet: Fig. 3. Effects of Thel on Wnt3a signaling proteins, AKT, and MAPKs. (A) Western blot analysis was used to detect Wnt3a, phospho-GSK3β (p-GSK3 β), and β-catenin in equal quantities. β -actin was detected in the same sample to normalize data obtained for lysates. (B, C) Western blot analysis was used to detect AKT, phospho-AKT (p-AKT) (B), phospho-ERK1 (p-ERK), ERK, phospho- JNK (p-JNK), JNK, phospho-p38 (p-p38), and p38 (C) in equal quantities. Total protein levels (AKT, ERK, JNK, and p38), respectively, were detected in the same sample to normalize data obtained for lysates. Representative data from three independent experiments are presented. Thel, thelephoric acid; MAPK, mitogen-activated protein kinase.

Article Snippet: Antibodies employed are listed as follows: Wnt3a (1:1000, #2721; Cell Signaling Technology, Beverly, MA, USA), p-GSK3 β (1:1000, #9336; Cell Signaling Technology), GSK3β (1:1000, #12456; Cell Signaling Technology), β-catenin (1:1000; Cell Signaling Technology), p-AKT (1:1000, #4060; Cell Signaling Technology), AKT (1:1000, #4691; Cell Signaling Technology), p-ERK1/2 (1:2000, #9101; Cell Signaling Technology), ERK1/2 (1:2000, #9102; Cell Signaling Technology), p-p38 (1:1000, #9211S, Cell Signaling Technology), p38 (1:1000, #9212, Cell Signaling Technology), p-JNK (1:500, #9251; Cell Signaling Technology), JNK (1:1000, #9252; Cell Signaling Technology), β-actin (C4) (1:1000, #sc-47778; Santa Cruz Biotechnology, Santa Cruz, CA, USA), BMP2 (1: 500, #CSB-PAO9419AORb; CUSABIO, Houston, TX, USA), RUNX2 (1:2000, #12556; Cell Signaling Technology), p-Smad1/5/8 (1:1000, #13820; Cell Signaling Technology), MMP13 (1:1000, #NBP1-45723SS; Novus Biologicals, Centennial, CO, USA).

Techniques: Western Blot

Journal: Molecular medicine reports

Article Title: MicroRNA‑214 targets Wnt3a to suppress liver cancer cell proliferation.

doi: 10.3892/mmr.2017.7483

Figure Lengend Snippet: Figure 1. miR‑214 is downregulated in liver cancer and targets Wnt3a. (A) Reverse transcription‑quantitative polymerase chain reaction was performed to examine the expression of miR‑214 in 24 paired human hepatocellular carcinoma and non‑tumor tissues. (B) Relative expression of miR‑214 in liver cancer cell lines and a normal liver cell line. **P<0.01; *P<0.05. (C) miR‑214 seed region sequence in the 3'UTR of Wnt3a. (D) Wnt3a protein expression as detected by immunohistochemistry. (E) Protein expression levels of Wnt3a were measured by western blot analysis in HepG2 cells transfected with miR‑214 or miR‑ctrl. (F) miR‑214 was co‑transfected with pmirGLO, pmirGLO‑Wnt3a‑3'‑UTR‑wt or pmirGLO‑Wnt3a‑3'‑UTR‑mut in HepG2 cells. Relative luciferase activity was measured after 48 h. *P<0.05 vs. control. miR, microRNA; mut/M, mutant; UTR, untranslated region; wt/W, wild‑type.

Article Snippet: Membranes were then incubated with rabbit anti-human Wnt3a (cat no. bs-1700R; 1:100; Beijing Biosynthesis Biotechnology Co., Ltd.) and rabbit anti-human GAPDH (cat no. 10494-1-AP; 1:2,000; ProteinTech Group, Inc., Chicago, IL, USA) at 4 ̊C overnight.

Techniques: Polymerase Chain Reaction, Expressing, Sequencing, Immunohistochemistry, Western Blot, Transfection, Luciferase, Activity Assay, Control, Mutagenesis

Journal: Molecular medicine reports

Article Title: MicroRNA‑214 targets Wnt3a to suppress liver cancer cell proliferation.

doi: 10.3892/mmr.2017.7483

Figure Lengend Snippet: Figure 2. miR‑214 inhibits the proliferation of liver cancer cells. CCK8 assay was performed to detect the effects of miR‑214 on cell proliferation at 24, 48, and 72 h in (A) HepG2 and (B) Hep3B cells. CCK8 assay was performed to detect the effects of siWnt3a on cell proliferation at 24, 48 and 72 h in (C) HepG2 and (D) Hep3B cells. Wnt3a overexpression vector was co‑transfected with miR‑ctrl or miR‑214 into (E) HepG2 and (F) Hep3B cells, and cell proliferation was detected by CCK8 assay. *P<0.05; **P<0.01 vs. miR‑ctrl + Wnt3a‑ctrl. CCK8, Cell Counting kit‑8; ctrl, control; miR, microRNA; OD, optical density; si, small interfering RNA.

Article Snippet: Membranes were then incubated with rabbit anti-human Wnt3a (cat no. bs-1700R; 1:100; Beijing Biosynthesis Biotechnology Co., Ltd.) and rabbit anti-human GAPDH (cat no. 10494-1-AP; 1:2,000; ProteinTech Group, Inc., Chicago, IL, USA) at 4 ̊C overnight.

Techniques: CCK-8 Assay, Over Expression, Plasmid Preparation, Control, Small Interfering RNA

Journal: Molecular medicine reports

Article Title: MicroRNA‑214 targets Wnt3a to suppress liver cancer cell proliferation.

doi: 10.3892/mmr.2017.7483

Figure Lengend Snippet: Figure 3. Overexpression of miR‑214 or Wnt3a silencing affects cell cycle progression. Cell cycle analysis of (A) HepG2 and (B) Hep3B cells following transfection with miR‑214 or miR‑ctrl for 48 h. Cell cycle analysis of (C) HepG2 and (D) Hep3B cells following transfection with siWnt3a or si‑ctrl for 48 h. *P<0.05. ctrl, control; miR, microRNA; si, small interfering RNA.

Article Snippet: Membranes were then incubated with rabbit anti-human Wnt3a (cat no. bs-1700R; 1:100; Beijing Biosynthesis Biotechnology Co., Ltd.) and rabbit anti-human GAPDH (cat no. 10494-1-AP; 1:2,000; ProteinTech Group, Inc., Chicago, IL, USA) at 4 ̊C overnight.

Techniques: Over Expression, Cell Cycle Assay, Transfection, Control, Small Interfering RNA