rabbit anti phospho β catenin thr41  (Bioss)

Journal: Frontiers in Oncology

Article Title: Retinoic Acid-Induced 2 (RAI2) Is a Novel Antagonist of Wnt/β-Catenin Signaling Pathway and Potential Biomarker of Chemosensitivity in Colorectal Cancer

doi: 10.3389/fonc.2022.805290

Figure Lengend Snippet: RAI2 suppressed activation of Wnt/β-catenin signaling pathway. (A) Dual-Luciferase reporter assay analyzed the effect of RAI2 plasmid transfection on wild-type β-catenin (β-catenin-wt)-induced activation of Wnt/β-catenin signaling in LoVo/HCT116 cells with or without LiCl/XAV939 addition (the concentrations of LiCl and XAV939 were 20mM and 10 µM respectively). LiCl and XAV939 were agonist and antagonist of Wnt/β-catenin signaling respectively. (B) Western blot analysis of RAI2, β-catenin, phosphorylated β-catenin (p-β-catenin) CtBP2 and the Wnt/β-catenin signaling target genes c-Myc, cylinD1, ASCL2, and LGR5 expression in LoVo/HCT116 cells with RAI2/empty vector transfection and RAI2-transfected cells with LiCl addition (20mM). The same experiment was also performed in shNC/shRAI2-transfected SW620 cells and shRAI2-transfected SW620 cells with XAV939 addition (10 µM). p-values: **<0.01; ***<0.001.

Article Snippet: Rabbit polyclonal antibody against RAI2 (Abcam, ST. Louis, MO) was diluted at 1:30, Rabbit anti-phospho-β-catenin (Thr41) (BIOSS, Beijing, China) was diluted at 1:1200, Rabbit anti- achaete-scute complex homolog 2 (ASCL2) (BIOSS, Beijing, China) was diluted at 1:200 and Rabbit anti-Leucine-rich repeat-containing G-protein coupled receptor 5 (LGR5) (Abcam, ST. Louis, MO) was diluted at 1:200.

Techniques: Activation Assay, Luciferase, Reporter Assay, Plasmid Preparation, Transfection, Western Blot, Expressing

Journal: Frontiers in Oncology

Article Title: Retinoic Acid-Induced 2 (RAI2) Is a Novel Antagonist of Wnt/β-Catenin Signaling Pathway and Potential Biomarker of Chemosensitivity in Colorectal Cancer

doi: 10.3389/fonc.2022.805290

Figure Lengend Snippet: RAI2 inhibits the nuclear translocation of β-catenin and suppresses the expression of Wnt signaling target genes. (A) Immunofluorescence analysis of expression and localization of RAI2 and β-catenin in LoVo cells with empty vector/RAI2/RAI2-M (mutant type RAI2 plasmid) transfection. The left panel showed the expression and localization of RAI2 in LoVo cells before and after RAI2/RAI2-M transfection; the right panel showed the expression and localization of β-catenin in LoVo cells before and after RAI2/RAI2-M transfection. (B) Western blot analysis of RAI2, β-catenin, phosphorylated β-catenin (p-β-catenin), CtBP2, and the Wnt/β-catenin signaling target genes c-Myc, CylinD1, ASCL2, and LGR5 expression in LoVo/HCT116 cells with empty vector/RAI2/RAI2-M (mutant type RAI2 plasmid) transfection and SW620 cells with shNC/shRAI2 transfection. (C) Coimmunoprecipitation (Co-IP) analysis of CtBP2 and APC interaction by anti-CtBP2 antibody using cell lysates from LoVo and HCT116 cells transfected with vector, RAI2 expression vector or mutant type RAI2 vector (RAI2-M). Extracted cell lysates of LoVo and HCT116 cells before adding antibodies were used for GAPDH detection (Input).

Article Snippet: Rabbit polyclonal antibody against RAI2 (Abcam, ST. Louis, MO) was diluted at 1:30, Rabbit anti-phospho-β-catenin (Thr41) (BIOSS, Beijing, China) was diluted at 1:1200, Rabbit anti- achaete-scute complex homolog 2 (ASCL2) (BIOSS, Beijing, China) was diluted at 1:200 and Rabbit anti-Leucine-rich repeat-containing G-protein coupled receptor 5 (LGR5) (Abcam, ST. Louis, MO) was diluted at 1:200.

Techniques: Translocation Assay, Expressing, Immunofluorescence, Plasmid Preparation, Mutagenesis, Transfection, Western Blot, Co-Immunoprecipitation Assay

Journal: Frontiers in Oncology

Article Title: Retinoic Acid-Induced 2 (RAI2) Is a Novel Antagonist of Wnt/β-Catenin Signaling Pathway and Potential Biomarker of Chemosensitivity in Colorectal Cancer

doi: 10.3389/fonc.2022.805290

Figure Lengend Snippet: Low expression of RAI2 was associated with a low level of phosphorylation of β-catenin and poor outcome in CRC. (A) Left panel: IHC staining of RAI2, phosphorylated β-catenin (p-β-catenin), ASCL2, and LGR5 in CRC tissues; right panel: the correlation between RAI2 expression and expression of p-β-catenin, ASCL2, and LGR5 (Spearman rank correlation test). (B) Kaplan-Meier curves show the association of the 5-year overall survival (OS) rate and relapse-free survival (RFS) rate of colorectal cancer patients with the expression status of RAI2. Black, RAI2-low expressed colorectal cancer patients (n = 101, log-rank test); red, RAI2-high expressed colorectal cancer patients (n = 197, log-rank test).

Article Snippet: Rabbit polyclonal antibody against RAI2 (Abcam, ST. Louis, MO) was diluted at 1:30, Rabbit anti-phospho-β-catenin (Thr41) (BIOSS, Beijing, China) was diluted at 1:1200, Rabbit anti- achaete-scute complex homolog 2 (ASCL2) (BIOSS, Beijing, China) was diluted at 1:200 and Rabbit anti-Leucine-rich repeat-containing G-protein coupled receptor 5 (LGR5) (Abcam, ST. Louis, MO) was diluted at 1:200.

Techniques: Expressing, Immunohistochemistry

Journal: Frontiers in Oncology

Article Title: Retinoic Acid-Induced 2 (RAI2) Is a Novel Antagonist of Wnt/β-Catenin Signaling Pathway and Potential Biomarker of Chemosensitivity in Colorectal Cancer

doi: 10.3389/fonc.2022.805290

Figure Lengend Snippet: Mechanisms model for the regulation of RAI2 in Wnt/β-catenin signaling. ① RAI2 inhibits the interaction of CtBP2 and APC. ② RAI2 suppresses the expression of CtBP2.

Article Snippet: Rabbit polyclonal antibody against RAI2 (Abcam, ST. Louis, MO) was diluted at 1:30, Rabbit anti-phospho-β-catenin (Thr41) (BIOSS, Beijing, China) was diluted at 1:1200, Rabbit anti- achaete-scute complex homolog 2 (ASCL2) (BIOSS, Beijing, China) was diluted at 1:200 and Rabbit anti-Leucine-rich repeat-containing G-protein coupled receptor 5 (LGR5) (Abcam, ST. Louis, MO) was diluted at 1:200.

Techniques: Expressing