wnt1 rabbit polyclonal antibody  (Proteintech)

Journal: Frontiers in Pharmacology

Article Title: Thymopentin-Mediated Inhibition of Cancer Stem Cell Stemness Enhances the Cytotoxic Effect of Oxaliplatin on Colon Cancer Cells

doi: 10.3389/fphar.2022.779715

Figure Lengend Snippet: Effect of TP5 on Wnt/β-catenin signaling pathway. (A–F) Typical Western blot analysis (A) and pooled data of protein expression levels of PI3K-P110 β (B) , WNT1 (C) , P-AKT (D) , P-β-catenin (E) and β-catenin (F) . Histograms are expressed as induction rates in triplicate for the GAPDH control. All data are expressed as mean ± SD; * p < 0.05, ** p < 0.01 and *** p < 0.001, one-way ANOVA with Dunnett’s post hoc test, (B) , F (4, 10) = 7.486, p = 0.0047, (C) , F (4, 10) = 11.47, p = 0.0009, (D) , F (4, 10) = 6.387, p = 0.0081, (E) , F (4, 10) = 7.421, p = 0.0048, (F) , F (4, 10) = 1.342, p = 0.3204; ns, not significant.

Article Snippet: The membranes were incubated overnight at 4°C with primary antibody, and secondary antibodies anti-rabbit IgG HRP-conjugated (1: 3,000, 7074S, Cell Signaling Technology) and anti-mouse IgG HRP-conjugated (1: 3,000, 7076S, Cell Signaling Technology) were incubated at 25°C for incubate for 2 h. The primary antibodies used are Beta Catenin Rabbit Polyclonal Antibody (1: 1,000, 51067-2-AP, Proteintech), WNT1 Rabbit Polyclonal Antibody (1: 1,000, 27935-1-AP, Proteintech), Phospho-beta Catenin (Ser33/37/Thr41) antibody (1: 1,000, 9561, Cell Signaling Technology), AKT1 (Phospho-Ser473 + Tyr474) antibody (1: 1,000, 12669, Abcam), PI3K p110 (beta) polyclonal antibody (1: 1,000, 20584-1-AP, Proteintech), AKT1 polyclonal antibody (1: 1,000, 10176-2-AP, Proteintech), and GAPDH (1: 10,000, 60004-I-Ig, Proteintech).

Techniques: Western Blot, Expressing, Control

Journal: Scientific Reports

Article Title: miRepress: modelling gene expression regulation by microRNA with non-conventional binding sites

doi: 10.1038/srep22334

Figure Lengend Snippet: ( a ) Predicted interaction of hsa-miR-367-5p and WNT1 3′ UTR sequence. miR-367-5p has no conventional 5′ end seed matched site in WNT1 3′ UTR, but it contains two non-conventional 3′ end matched binding site in the positions 1566-1575 and 2035-2040 in the WNT1 mRNA 3′ UTR sequence. ( b ) MCF-7 cells were co-transfected with mimic of hsa-367-5p miRNA or non-specific control RNA oligo (Nm) and plasmid DNA encoding wild-type luciferase gene or mutant luciferase gene. Note that wild-type, not the mutant, luciferase gene was inhibited by hsa-367-3p miRNA. The luminescence signal in wild-type luciferase was reduced by approximately 71%. ( c ) Predicted interaction of hsa-miR-367-5p and luciferase coding sequence. miR-367-5p has no conventional 5′ end seed matched site in luciferase gene, but it contains one non-conventional 3′ end matched binding site in the 978–984 region in the luciferase gene coding sequence. ( d ) The predicted binding site of hsa-miR-367-5p on firefly luciferase coding sequence is mutated (GGUUGCA- > GGUCGC) so that hsa-miR-367-5p could no longer bind and repress luciferase gene. ( e ) MCF-7 cells were co-transfected with mimic hsa-miR-367-5p miRNA or inhibitor of hsa-miR-367-5p or mutant hsa-miR-367-5p or non-specific control RNA oligo and plasmid DNA encoding mutant luciferase gene tagged with 3′-UTR of WNT1. The luminescence signal was reduced approximately 79% in case of co-transfection of mimic has-miR-367-5p. Note that luciferase signal was reduced in the presence of hsa-367-5p miRNA and that ~30% luciferase was recovered in presence of has-miR-367-5p inhibitor or mutant hsa-miR-367-5p. *p < 0.05, Nm vs hsa-miR-367-5p. ( f ) hsa-miR-367-5p is mutated in its predicted binding site in its 3′ end i.e. base 14–19 (UGCAAC- > UCACCC) so that the binding of mutant hsa-miR-367-5p 3′ end with the two sites in WNT1 3′ UTR is perturbed.

Article Snippet: The membranes were incubated with rabbit polyclonal NMHC II-B (1:2000, Sigma), rabbit polyclonal WNT1 (1:1000, Santa Cruz) or mouse polyclonal GAPDH (1:4000, Santa Cruz) at 4 °C for overnight.

Techniques: Sequencing, Binding Assay, Transfection, Control, Plasmid Preparation, Luciferase, Mutagenesis, Cotransfection

Journal: Experimental and Therapeutic Medicine

Article Title: Immunohistochemical demonstration of alteration of β-catenin during tumor metastasis by different mechanisms according to histology in lung cancer

doi: 10.3892/etm.2014.2095

Figure Lengend Snippet: Immunohistochemical changes from primary to metastatic tumors.

Article Snippet: The sections were incubated with rabbit polyclonal antibody for Wnt1 (H-89; Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA; dilution 1:100), mouse monoclonal antibody for β-catenin (5H10; Zymed Laboratories, Invitrogen Life Technologies, Carlsbad, CA, USA; dilution 1:1,000) and mouse monoclonal antibody for E-cadherin (SPM471; Thermo Fisher Scientific, Waltham, MA, USA; dilution 1:150) at 4°C overnight.

Techniques: Immunohistochemical staining

Journal: Experimental and Therapeutic Medicine

Article Title: Immunohistochemical demonstration of alteration of β-catenin during tumor metastasis by different mechanisms according to histology in lung cancer

doi: 10.3892/etm.2014.2095

Figure Lengend Snippet: (A) The intact membrane expression of β-catenin in the primary tumor; (B) the alteration of β-catenin in the corresponding metastatic tumor; (C) no expression of Wnt1 in the primary tumor; (D) Wnt1 overexpression in the corresponding metastatic tumor; (E) the intact membrane expression of E-cadherin in the primary tumor and; (F) the membrane loss of E-cadherin in the corresponding metastatic tumor.

Article Snippet: The sections were incubated with rabbit polyclonal antibody for Wnt1 (H-89; Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA; dilution 1:100), mouse monoclonal antibody for β-catenin (5H10; Zymed Laboratories, Invitrogen Life Technologies, Carlsbad, CA, USA; dilution 1:1,000) and mouse monoclonal antibody for E-cadherin (SPM471; Thermo Fisher Scientific, Waltham, MA, USA; dilution 1:150) at 4°C overnight.

Techniques: Membrane, Expressing, Over Expression

Journal: Experimental and Therapeutic Medicine

Article Title: Immunohistochemical demonstration of alteration of β-catenin during tumor metastasis by different mechanisms according to histology in lung cancer

doi: 10.3892/etm.2014.2095

Figure Lengend Snippet: Comparison of β-catenin, E-cadherin and Wnt1 expression between primary and metastatic tumors.

Article Snippet: The sections were incubated with rabbit polyclonal antibody for Wnt1 (H-89; Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA; dilution 1:100), mouse monoclonal antibody for β-catenin (5H10; Zymed Laboratories, Invitrogen Life Technologies, Carlsbad, CA, USA; dilution 1:1,000) and mouse monoclonal antibody for E-cadherin (SPM471; Thermo Fisher Scientific, Waltham, MA, USA; dilution 1:150) at 4°C overnight.

Techniques: Comparison, Expressing, Over Expression

Journal: Experimental and Therapeutic Medicine

Article Title: Immunohistochemical demonstration of alteration of β-catenin during tumor metastasis by different mechanisms according to histology in lung cancer

doi: 10.3892/etm.2014.2095

Figure Lengend Snippet: Comparison of β-catenin vs. E-cadherin and Wnt1 expression in adenocarcinomas.

Article Snippet: The sections were incubated with rabbit polyclonal antibody for Wnt1 (H-89; Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA; dilution 1:100), mouse monoclonal antibody for β-catenin (5H10; Zymed Laboratories, Invitrogen Life Technologies, Carlsbad, CA, USA; dilution 1:1,000) and mouse monoclonal antibody for E-cadherin (SPM471; Thermo Fisher Scientific, Waltham, MA, USA; dilution 1:150) at 4°C overnight.

Techniques: Comparison, Expressing