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

Journal: Oncology reports

Article Title: Matrine induces mitochondrial apoptosis in cisplatin-resistant non-small cell lung cancer cells via suppression of β-catenin/survivin signaling.

doi: 10.3892/or.2015.3844

Figure Lengend Snippet: Figure 2. Increased β-catenin activity in the CDDP-resistant NSCLC cells. (A) Western blot analysis of total and non-phospho (active) β-catenin (non- p-β-cat). Representative blots are shown in the top panels, and densitometric quantification of the blots is shown in the bottom graph. (B) CDDP-resistant A549 and H460 cells and their parental cells were transfected with pTop- Flash and pRL-TK. After incubation for 24 h, the cells were treated with matrine for an additional 48 h. The cells were collected and tested for luciferase activities. The results are expressed as relative luciferase activity (RLA) compared to the parental cells arbitrarily assigned as 1. *P<0.05 vs. the untreated cells. CDDP, cisplatin; NSCLC, non-small cell lung cancer.

Article Snippet: Primary antibodies used in the present study are as follows: anti-β-catenin (#9562), anti-phosphoβ-catenin (Ser33/37/Thr41) (#9561), anti-non-phospho (active) β-catenin (Ser33/37/Thr41) (#4270), anti-GSK-3β (#9315), anti-phospho-GSK-3β (Ser9) (#9323) (Cell Signaling Technology, Danvers, MA, uSA), anti-survivin (sc-10811) and anti-β-actin (sc-130301) (Santa Cruz Biotechnology, Santa Cruz, CA, USA).

Techniques: Activity Assay, Western Blot, Transfection, Incubation, Luciferase

Journal: Oncology reports

Article Title: Matrine induces mitochondrial apoptosis in cisplatin-resistant non-small cell lung cancer cells via suppression of β-catenin/survivin signaling.

doi: 10.3892/or.2015.3844

Figure Lengend Snippet: Figure 3. Effect of matrine treatment on β-catenin signaling in CDDP-resistant NSCLC cells. CDDP-resistant (A) A549 and (B) H460 cells were untreated or treated with 1 or 2 g/l of matrine for 48 h, and western blot analysis was carried out to analyze changes in β-catenin signaling. Representative blots are shown. non-p-β-Cat, non-phospho β-catenin. (C) CDDP-resistant A549 and H460 cells were pre-transfected with pTopFlash and pRL-TK and 24 h later, treated with or without matrine for an additional 48 h. The results are expressed as relative luciferase activity (RLA) compared to the control cells (without matrine treat- ment) arbitrarily assigned as 100%. *P<0.05 vs. the control cells. (D) Western blot analysis of survivin expression in the CDDP-resistant NSCLC cells with or without matrine treatment. CDDP, cisplatin; NSCLC, non-small cell lung cancer.

Article Snippet: Primary antibodies used in the present study are as follows: anti-β-catenin (#9562), anti-phosphoβ-catenin (Ser33/37/Thr41) (#9561), anti-non-phospho (active) β-catenin (Ser33/37/Thr41) (#4270), anti-GSK-3β (#9315), anti-phospho-GSK-3β (Ser9) (#9323) (Cell Signaling Technology, Danvers, MA, uSA), anti-survivin (sc-10811) and anti-β-actin (sc-130301) (Santa Cruz Biotechnology, Santa Cruz, CA, USA).

Techniques: Western Blot, Transfection, Luciferase, Activity Assay, Control, Expressing

Journal: The Journal of cell biology

Article Title: The transition zone protein Rpgrip1l regulates proteasomal activity at the primary cilium.

doi: 10.1083/jcb.201408060

Figure Lengend Snippet: Figure 4. Rpgrip1l deficiency causes impaired proteasomal activity at primary cilia. (A–F and I–L) MEFs were isolated from E12.5 WT and Rpgrip1l−/− embryos. (A and B) Western blot analysis of WT and Rpgrip1l−/− MEF lysates (n = 4 embryos, respectively). (C) Western blot analysis of WT and Rpgrip1l−/− MEF lysates (n = 3 embryos, respectively). (A–C) Actin serves as a loading control. (A) Phospho-(S33/37/T41)-β-Catenin is significantly increased in serum-starved Rpgrip1l−/− MEF lysates (82% of all cells had cilia; in serum-starved Rpgrip1l+/+ MEF lysates, 88.67% of all cells possessed cilia) but not in non–serum-starved Rpgrip1l−/− MEF lysates (4% of all cells displayed cilia; in non–serum-starved Rpgrip1l+/+ MEF lysates, 6.67% of all cells carried cilia; C). (B) Non–phospho-(S33/37/T41)-β-Catenin is unaltered in serum-starved Rpgrip1l−/− MEF lysates. Black lines indicate that interven- ing lanes have been spliced out. (D–F, I, and L) Immunofluorescence on MEFs of E12.5 WT and Rpgrip1l−/− embryos (both genotypes: p-β-Catenin: n = 5; p-β-Catenin (3D-SIM, n = 3; Ubiquitin, n = 4; Gli3-190, n = 6; ZsProSensor-1, n = 3; n refers to the number of embryos, respectively). Per embryo, 15 cilia were quantified for p-β-Catenin, 10 cilia were quantified for p-β-Catenin (3D-SIM) and for Ubiquitin, and 20 cilia were quantified for Gli3-190. (G and H) Immunofluorescence on limbs of E12.5 WT and Rpgrip1l−/− embryos (n = 3 embryos, respectively). Per embryo, 20 cilia were quantified for p-β-Catenin and Ubiquitin. All quantified proteins are shown in red (D–J), the ciliary axoneme is marked by acetylated α-tubulin (green; D–J), and the BB is marked by γ-tubulin (blue; D–F, H, and I) or by Pcnt2 (blue; G). (J and K) Immunofluorescence on MEFs of WT embryos (n = 4). 25 cilia per embryo were used for phospho-(S33/37/T41)-β-Catenin and cilia length quantification. (L) Proteasome activity assay on WT and Rpgrip1l−/− MEFs. Cilia are marked by acetylated α-tubulin (α-Tub), and centrosomes/basal bodies are marked by γ-tubulin. Colored squares mark cilia with basal bodies (yellow squares) as well as centrosomes (red squares), which are presented magnified. The green ZsProSensor-1 protein signal is exclusively detected at the ciliary base in Rpgrip1l−/− MEFs. Error bars show standard error of the mean. *, P < 0.05; **, P < 0.01; ***, P < 0.001. Bars, 1 µm.

Article Snippet: We used the following primary antibodies: rabbit anti-actin (A2066; Sigma-Aldrich), mouse anti-FLAG (200472; Agilent Technologies), mouse anti-Gapdh (G8795; Sigma-Aldrich), goat anti-Gli3 (AF3690; R&D systems), mouse anti-HA (MMS-101P; Covance), rabbit anti-Hprt (ab10479; Abcam), rabbit anti-Myc (sc-789; Santa Cruz Biotechnology, Inc.), rabbit anti-phospho-(S33/37/T41)-β-Catenin (9561; Cell Signaling Technology), rabbit anti–nonphospho-(S33/37/ T41)-β-Catenin (4270; Cell Signaling Technology), rabbit anti-Psma5 (PA1-1962; Thermo Fisher Scientific), mouse anti-Pcnt (611814; BD), goat anti-Pcnt2 (sc-28145; Santa Cruz Biotechnology, Inc.), goat anti-Psmd2 (IMG-3751; Imgenex), rabbit anti-Psmd3 (S2824; Sigma-Aldrich), rabbit anti-Psmd4 (14899-1-AP; Proteintech), rabbit anti-Smo (ab72130; Abcam), mouse anti–acetylated α-tubulin (T6793; Sigma-Aldrich), mouse anti–γ-tubulin (T6557; Sigma-Aldrich), and rabbit anti-Ubiquitin (U5379; Sigma-Aldrich).

Techniques: Activity Assay, Isolation, Western Blot, Control, Immunofluorescence, Ubiquitin Proteomics

Journal: The Journal of cell biology

Article Title: The transition zone protein Rpgrip1l regulates proteasomal activity at the primary cilium.

doi: 10.1083/jcb.201408060

Figure Lengend Snippet: Figure 8. Proteasomal activity is unaltered at Rpgrip1l−/− centrosomes. (A–C) Immunofluorescence on MEFs isolated from E12.5 WT and Rpgrip1l−/− embryos (WT: p-β-Catenin (treated with DMSO or MG132): n = 3 embryos; both genotypes: p-β-Catenin and Ubiquitin: n = 3 embryos, respectively). The ciliary axoneme is marked by acetylated α-tubulin (green) and the centrosomes (basal bodies in case of ciliary presence) by γ-tubulin (blue). All quantified proteins are shown in red. An axonemal-like green staining is not visible, demonstrating that the blue staining marks centrosomes. (A) After treatment of WT MEFs with the proteasome inhibitor MG132, the amount of phospho-(S33/37/T41)-β-Catenin is significantly increased at the centrosome. (B and C) The amounts of phospho-(S33/37/T41)-β-Catenin and Ubiquitin are unaltered at the centrosome of Rpgrip1l−/− MEFs. (A–C) Per embryo, 20 cilia were used in the quantifications. Error bars show standard error of the mean. *, P < 0.05. Bars, 0.5 µm.

Article Snippet: We used the following primary antibodies: rabbit anti-actin (A2066; Sigma-Aldrich), mouse anti-FLAG (200472; Agilent Technologies), mouse anti-Gapdh (G8795; Sigma-Aldrich), goat anti-Gli3 (AF3690; R&D systems), mouse anti-HA (MMS-101P; Covance), rabbit anti-Hprt (ab10479; Abcam), rabbit anti-Myc (sc-789; Santa Cruz Biotechnology, Inc.), rabbit anti-phospho-(S33/37/T41)-β-Catenin (9561; Cell Signaling Technology), rabbit anti–nonphospho-(S33/37/ T41)-β-Catenin (4270; Cell Signaling Technology), rabbit anti-Psma5 (PA1-1962; Thermo Fisher Scientific), mouse anti-Pcnt (611814; BD), goat anti-Pcnt2 (sc-28145; Santa Cruz Biotechnology, Inc.), goat anti-Psmd2 (IMG-3751; Imgenex), rabbit anti-Psmd3 (S2824; Sigma-Aldrich), rabbit anti-Psmd4 (14899-1-AP; Proteintech), rabbit anti-Smo (ab72130; Abcam), mouse anti–acetylated α-tubulin (T6793; Sigma-Aldrich), mouse anti–γ-tubulin (T6557; Sigma-Aldrich), and rabbit anti-Ubiquitin (U5379; Sigma-Aldrich).

Techniques: Activity Assay, Immunofluorescence, Isolation, Ubiquitin Proteomics, Staining

Journal: Human molecular genetics

Article Title: Haploinsufficiency of RCBTB1 is associated with Coats disease and familial exudative vitreoretinopathy.

doi: 10.1093/hmg/ddw041

Figure Lengend Snippet: Figure 4. Knockdown of RCBTB1 significantly reduced the activation of the Norrin- or Wnt3a-mediated Wnt/β-catenin pathway by reducing the nuclear accumulation of β-

Article Snippet: Antibodies to RCBTB1 (ab154649, Abcam®), active (nonphospho) β-catenin (#8814, Cell Signaling Technology®), Lamin B1 (ab133741, Abcam®), GAPDH (NB300-322, Novus Biologicals) and β-actin (8H10D10, Cell Signaling Technology®) were used to detect and quantify the proteins.

Techniques: Knockdown, Activation Assay

Journal: PLoS ONE

Article Title: The Wnt Serpentine Receptor Frizzled-9 Regulates New Bone Formation in Fracture Healing

doi: 10.1371/journal.pone.0084232

Figure Lengend Snippet: WT: upper panel, Fdz9 −/− : lower panel. β-catenin was expressed in osteoblasts (OB) and proliferating chondroblasts (CB) but to a lesser extend in hypertrophic chondrocytes (HC). C: cortex. There were no differences between both genotypes. Only TRAP positive cells with ≥2 nuclei were identified as osteoclasts (OC). TRAP-staining either revealed no significant differences between both genotypes.

Article Snippet: Immunostaining of β-catenin, Runx2, Osteocalcin, and chemokines Cxcl5 and Ccl2 were performed on paraffin-embedded sections from day 10 using a polyclonal anti-rabbit β-catenin antibody (EMD Millipore Corporation, Massachusetts, USA), anti-Runx2 antibody (Cell Signaling Technology Inc., Danvers, USA), anti-Osteocalcin antibody (Biorbyt Ltd., Cambridge, UK), anti-Cxcl5 antibody (Bioss Inc., Woburn, USA), and anti-Ccl2 antibody (Bioss Inc., Woburn, USA) respectively.

Techniques: Staining

Journal: Frontiers in Cell and Developmental Biology

Article Title: Signaling Proteins Recruited to the Sperm Binding Site: Role of β-Catenin and Rho A

doi: 10.3389/fcell.2022.886664

Figure Lengend Snippet: Localization of β-catenin at the sperm binding site.Oocytes fixed at different stages of fertilization were labeled with anti- β-catenin (green) and Alexa Fluor-568 phalloidin (red) as well as Hoechst 33342 (blue). (A) Recently bound sperm. (B) bound sperm with microvilli exhibiting β-catenin labelling. (C) β-catenin plaque colocalizing with the actin layer. (D) sperm undergoing incorporation into the ooplasm with a single bright patch of β-catenin remaining. Images were selected from 30 oocytes recovered from 4 females. Magnification is indicated by the bar.

Article Snippet: The specificity of the non-phospho (Active) β−catenin (Ser33/37/Thr41) antibody was tested by addition of a synthetic peptide (DSGIHSGATTT aa32-aa42) from GenScript (Piscataway NJ) as a competitive inhibitor of the primary antibody at 1:100 excess ( ).

Techniques: Binding Assay, Labeling

Journal: Frontiers in Cell and Developmental Biology

Article Title: Signaling Proteins Recruited to the Sperm Binding Site: Role of β-Catenin and Rho A

doi: 10.3389/fcell.2022.886664

Figure Lengend Snippet: Distribution of f-actin and β-catenin in the oocyte cortex.MII oocyte from a virgin female was treated by prolonged fixation and labeled with anti- β-catenin (green) and Alexa Fluor-568 phalloidin (red) to label f-actin, as well as Hoechst 33342 (blue). Z-stack images viewed in volume mode (A) , a tangential image (B) , and an equatorial image (C) reveal fenestrae appearing as holes in the cortical actin layer (A,B) or as regions of thinner f-actin in the equatorial view (C) . Images were selected from 10 oocytes from three females. Magnification is indicated by the bar.

Article Snippet: The specificity of the non-phospho (Active) β−catenin (Ser33/37/Thr41) antibody was tested by addition of a synthetic peptide (DSGIHSGATTT aa32-aa42) from GenScript (Piscataway NJ) as a competitive inhibitor of the primary antibody at 1:100 excess ( ).

Techniques: Labeling

Journal: Cell Proliferation

Article Title: Fam70A binds Wnt5a to regulate meiosis and quality of mouse oocytes

doi: 10.1111/cpr.12825

Figure Lengend Snippet: Fam70A and Wnt5a regulate β‐catenin activity. A, Co‐IP showed that Fam70A interacted with β‐catenin within oocytes. B, Co‐IP showed that Wnt5a interacted with β‐catenin within oocytes. C, Western blot showed that Fam70A depletion remarkably increased p‐β‐catenin level. D, Quantification of (C). E, Immunofluorescence showed that Fam70A depletion remarkably increased p‐β‐catenin at spindle poles. F, Western blot showed that Wnt5a knockdown remarkably increased p‐β‐catenin level. G, Quantification of (F). H, Western blot showed that APC knockdown remarkably decreased p‐β‐catenin level. I, Quantification of (H). Scale bar, 5 µm. * P < .05

Article Snippet: The following primary Abs were used: Rabbit anti‐Fam70a Ab (Cat#: bs‐11006R; Bioss); Rabbit anti‐Wnt5a Ab (Cat#: A12744; ABclonal); Rabbit anti‐adenomatous polyposis coli (APC) Ab (Cat#: AF0547; Affinity Biosciences); Rabbit anti‐Phospho Catenin Beta (Ser33/37/Thr41) Ab (Cat#: DF2989; Affinity Biosciences); Rabbit anti‐Phospho Akt (Thr308) Ab (Cat#: 13038; Cell Signaling Technology); Rabbit anti‐Phospho Akt (Ser473) Ab (Cat#: A5030; Bimake.Com); Mouse anti‐β‐actin Ab (Cat#: A5316‐100); Mouse anti‐β‐tubulin Ab (Cat#: sc‐5274); Mouse anti‐GAPDH Ab (Cat#: 30201ES60; YEASEN); Rabbit anti‐Beta‐Catenin Ab (Cat#: A5038; Bimake.Com); Mouse anti‐Strep II Tag Ab (Cat#: YFMA0054; YI FEI XUE BIOTECHNOLOGY); Mouse anti‐Flag Tag Ab (Cat#: YFMA0036; YI FEI XUE BIOTECHNOLOGY).

Techniques: Activity Assay, Co-Immunoprecipitation Assay, Western Blot, Immunofluorescence

Journal: Cell Proliferation

Article Title: Fam70A binds Wnt5a to regulate meiosis and quality of mouse oocytes

doi: 10.1111/cpr.12825

Figure Lengend Snippet: Fam70A working model. At cell membrane, Fam70A directly binds Wnt5a, which results in the inhibition of APC and activation of Akt (p‐Akt). APC promotes the phosphorylation (inactive form) of β‐catenin and stabilizes microtubules as well. β‐Catenin (active form) can get into the nucleus to promote gene transcription (but not in fully grown oocytes). Proper microtubule stability and p‐Akt together promote meiosis and quality of mouse oocytes

Article Snippet: The following primary Abs were used: Rabbit anti‐Fam70a Ab (Cat#: bs‐11006R; Bioss); Rabbit anti‐Wnt5a Ab (Cat#: A12744; ABclonal); Rabbit anti‐adenomatous polyposis coli (APC) Ab (Cat#: AF0547; Affinity Biosciences); Rabbit anti‐Phospho Catenin Beta (Ser33/37/Thr41) Ab (Cat#: DF2989; Affinity Biosciences); Rabbit anti‐Phospho Akt (Thr308) Ab (Cat#: 13038; Cell Signaling Technology); Rabbit anti‐Phospho Akt (Ser473) Ab (Cat#: A5030; Bimake.Com); Mouse anti‐β‐actin Ab (Cat#: A5316‐100); Mouse anti‐β‐tubulin Ab (Cat#: sc‐5274); Mouse anti‐GAPDH Ab (Cat#: 30201ES60; YEASEN); Rabbit anti‐Beta‐Catenin Ab (Cat#: A5038; Bimake.Com); Mouse anti‐Strep II Tag Ab (Cat#: YFMA0054; YI FEI XUE BIOTECHNOLOGY); Mouse anti‐Flag Tag Ab (Cat#: YFMA0036; YI FEI XUE BIOTECHNOLOGY).

Techniques: Membrane, Inhibition, Activation Assay