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Addgene inc active wnt2b v5 encoding homo wnt2b

HCC-TCM promotes M2 polarization via <t>Wnt2b/β-catenin</t> signalling. a THP-1 derived macrophages (THP-1-M) were incubated with 50% HCC-TCM for 48 h to obtain the HCC-educated macrophages (HCC-TAMs). Transcript expression levels of Wnt2b were determined in HCC-TAMs by qPCR. b The expression levels of Wnt2b (red) in CD68 + macrophages (green) were determined by immunofluorescence using TMA containing pairs of tumors and matched para-carcinoma tissues of HCC patients. c , d THP-1-M were transfected with control vectors or Wnt2B-V5 (over-Wnt2b) vectors for 48 h. The expression levels of CD163 and markers for M1 or M2 macrophages on/in these cells were determined by flow cytometry and qPCR, respectively. THP-1-M infected with control vectors, sh-Wnt2b or sh-CTNNB1 (β-catenin) vector were acquired as described in the Materials and Methods, and then incubated with 50% HCC-TCM for 48 h. The expression levels of CD163 and markers for M1 or M2 macrophages on/in these cells were determined by flow cytometry ( e , i ) and qPCR ( f , j ), respectively. The expression levels of β-catenin in HCC-TAMs that were infected with control vectors or sh-Wnt2b vectors were determined by western blotting and immunofluorescence respectively ( g , h ). One representative of at least three independent experiments is shown. qPCR, quantitative real-time PCR; HCC, hepatocellular carcinoma; TCM, tumour condition culture medium; TAMs, tumour-associated macrophages; 7721, SMMC-7721; TMA, Tissue microarray. Data are presented as mean ± SEM from at least three independent experiments (* p < 0.05, ** p < 0.01 and *** p < 0.001)

Active Wnt2b V5 Encoding Homo Wnt2b, supplied by Addgene inc, used in various techniques. Bioz Stars score: 92/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/active+wnt2b+v5+encoding+homo+wnt2b/pmc07788901-80-0-7?v=Addgene+inc
Average 92 stars, based on 2 article reviews

active wnt2b v5 encoding homo wnt2b - by Bioz Stars, 2026-07

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1) Product Images from "Promotion of epithelial-mesenchymal transformation by hepatocellular carcinoma-educated macrophages through Wnt2b/β-catenin/c-Myc signaling and reprogramming glycolysis"

Article Title: Promotion of epithelial-mesenchymal transformation by hepatocellular carcinoma-educated macrophages through Wnt2b/β-catenin/c-Myc signaling and reprogramming glycolysis

Journal: Journal of Experimental & Clinical Cancer Research : CR

doi: 10.1186/s13046-020-01808-3

HCC-TCM promotes M2 polarization via Wnt2b/β-catenin signalling. a THP-1 derived macrophages (THP-1-M) were incubated with 50% HCC-TCM for 48 h to obtain the HCC-educated macrophages (HCC-TAMs). Transcript expression levels of Wnt2b were determined in HCC-TAMs by qPCR. b The expression levels of Wnt2b (red) in CD68 + macrophages (green) were determined by immunofluorescence using TMA containing pairs of tumors and matched para-carcinoma tissues of HCC patients. c , d THP-1-M were transfected with control vectors or Wnt2B-V5 (over-Wnt2b) vectors for 48 h. The expression levels of CD163 and markers for M1 or M2 macrophages on/in these cells were determined by flow cytometry and qPCR, respectively. THP-1-M infected with control vectors, sh-Wnt2b or sh-CTNNB1 (β-catenin) vector were acquired as described in the Materials and Methods, and then incubated with 50% HCC-TCM for 48 h. The expression levels of CD163 and markers for M1 or M2 macrophages on/in these cells were determined by flow cytometry ( e , i ) and qPCR ( f , j ), respectively. The expression levels of β-catenin in HCC-TAMs that were infected with control vectors or sh-Wnt2b vectors were determined by western blotting and immunofluorescence respectively ( g , h ). One representative of at least three independent experiments is shown. qPCR, quantitative real-time PCR; HCC, hepatocellular carcinoma; TCM, tumour condition culture medium; TAMs, tumour-associated macrophages; 7721, SMMC-7721; TMA, Tissue microarray. Data are presented as mean ± SEM from at least three independent experiments (* p < 0.05, ** p < 0.01 and *** p < 0.001)

Figure Legend Snippet: HCC-TCM promotes M2 polarization via Wnt2b/β-catenin signalling. a THP-1 derived macrophages (THP-1-M) were incubated with 50% HCC-TCM for 48 h to obtain the HCC-educated macrophages (HCC-TAMs). Transcript expression levels of Wnt2b were determined in HCC-TAMs by qPCR. b The expression levels of Wnt2b (red) in CD68 + macrophages (green) were determined by immunofluorescence using TMA containing pairs of tumors and matched para-carcinoma tissues of HCC patients. c , d THP-1-M were transfected with control vectors or Wnt2B-V5 (over-Wnt2b) vectors for 48 h. The expression levels of CD163 and markers for M1 or M2 macrophages on/in these cells were determined by flow cytometry and qPCR, respectively. THP-1-M infected with control vectors, sh-Wnt2b or sh-CTNNB1 (β-catenin) vector were acquired as described in the Materials and Methods, and then incubated with 50% HCC-TCM for 48 h. The expression levels of CD163 and markers for M1 or M2 macrophages on/in these cells were determined by flow cytometry ( e , i ) and qPCR ( f , j ), respectively. The expression levels of β-catenin in HCC-TAMs that were infected with control vectors or sh-Wnt2b vectors were determined by western blotting and immunofluorescence respectively ( g , h ). One representative of at least three independent experiments is shown. qPCR, quantitative real-time PCR; HCC, hepatocellular carcinoma; TCM, tumour condition culture medium; TAMs, tumour-associated macrophages; 7721, SMMC-7721; TMA, Tissue microarray. Data are presented as mean ± SEM from at least three independent experiments (* p < 0.05, ** p < 0.01 and *** p < 0.001)

Techniques Used: Derivative Assay, Incubation, Expressing, Immunofluorescence, Transfection, Control, Flow Cytometry, Infection, Plasmid Preparation, Western Blot, Real-time Polymerase Chain Reaction, Microarray

The activation of Wnt2b/β-catenin signalling enhances TAMs-induced tumour-promoting effects. a THP-1 derived macrophages (THP-1-M) were transfected with control vectors or Wnt2B-V5 (over-Wnt2b) vectors for 48 h. These macrophages were incubated with RPMI 1640 for an additional 24 h to obtain the condition medium (CM). HCC cells were cultured in the presence of indicated CM for 48 h. The expression levels of EMT markers were determined by western blotting. ( b-e ) THP-1-M infected with control vectors, sh-Wnt2b or sh-CTNNB1 (β-catenin) vectors were acquired as described in Materials and Methods. These macrophages were incubated with 50% HCC-TCM for 48 h for the preparation of the different TAMs. These TAMs were incubated with RPMI 1640 for another 24 h to obtain the CM. b , c HCC cells were cultured in the presence of the indicated CM for 48 h. The expression levels of EMT markers were determined by western blotting. d HCC cells were incubated with culture medium (Ctrl) or the indicated CM for 24 h. The cell viability of each group was detected by MTT assay. e HCC cells were scratched with a plastic pipette tip and incubated with culture medium (Ctrl) or indicated CM for 24 h. The results of this wound healing assay were photographed and measured. One representative of at least three independent experiments is shown. qPCR, quantitative real-time PCR; HCC, hepatocellular carcinoma; TCM, tumour condition culture medium; TAMs, tumour-associated macrophages; 7721, SMMC-7721. Data are presented as mean ± SEM from at least three independent experiments (* p < 0.05, ** p < 0.01 and *** p < 0.001)

Figure Legend Snippet: The activation of Wnt2b/β-catenin signalling enhances TAMs-induced tumour-promoting effects. a THP-1 derived macrophages (THP-1-M) were transfected with control vectors or Wnt2B-V5 (over-Wnt2b) vectors for 48 h. These macrophages were incubated with RPMI 1640 for an additional 24 h to obtain the condition medium (CM). HCC cells were cultured in the presence of indicated CM for 48 h. The expression levels of EMT markers were determined by western blotting. ( b-e ) THP-1-M infected with control vectors, sh-Wnt2b or sh-CTNNB1 (β-catenin) vectors were acquired as described in Materials and Methods. These macrophages were incubated with 50% HCC-TCM for 48 h for the preparation of the different TAMs. These TAMs were incubated with RPMI 1640 for another 24 h to obtain the CM. b , c HCC cells were cultured in the presence of the indicated CM for 48 h. The expression levels of EMT markers were determined by western blotting. d HCC cells were incubated with culture medium (Ctrl) or the indicated CM for 24 h. The cell viability of each group was detected by MTT assay. e HCC cells were scratched with a plastic pipette tip and incubated with culture medium (Ctrl) or indicated CM for 24 h. The results of this wound healing assay were photographed and measured. One representative of at least three independent experiments is shown. qPCR, quantitative real-time PCR; HCC, hepatocellular carcinoma; TCM, tumour condition culture medium; TAMs, tumour-associated macrophages; 7721, SMMC-7721. Data are presented as mean ± SEM from at least three independent experiments (* p < 0.05, ** p < 0.01 and *** p < 0.001)

Techniques Used: Activation Assay, Derivative Assay, Transfection, Control, Incubation, Cell Culture, Expressing, Western Blot, Infection, MTT Assay, Transferring, Wound Healing Assay, Real-time Polymerase Chain Reaction

The inhibition of Wnt2b/β-catenin signaling reduces the tumour-promoting effect of HCC-TAMs in vivo. SMMC-7721 cells (6 × 10 6 ) with or without the indicated TAMs (1.5 × 10 6 ) were mixed with Matrigel (at ratio 4:1) and subcutaneously injected into the right subaxillary of 6-week old immunodeficient mice ( n = 6 mice/group). a , d Representative images of the subcutaneous tumors from each group. b , e Growth of subcutaneous tumours (left); the average tumour weight of each group at the time of euthanisation(right). c , f Representative images of immunohistochemistry staining of vimentin, E-cadherin, β-catenin, c-Myc in tumour tissues. g Overall survival HCC patients related to indicated gene expression levels, were generated by the GEPIA2 database via Kaplan–Meier analysis. HCC, hepatocellular carcinoma; TAMs, tumour-associated macrophages; IHC, Immunohistochemistry. Data are presented as mean ± SEM (* p < 0.05, ** p < 0.01 and *** p < 0.001)

Figure Legend Snippet: The inhibition of Wnt2b/β-catenin signaling reduces the tumour-promoting effect of HCC-TAMs in vivo. SMMC-7721 cells (6 × 10 6 ) with or without the indicated TAMs (1.5 × 10 6 ) were mixed with Matrigel (at ratio 4:1) and subcutaneously injected into the right subaxillary of 6-week old immunodeficient mice ( n = 6 mice/group). a , d Representative images of the subcutaneous tumors from each group. b , e Growth of subcutaneous tumours (left); the average tumour weight of each group at the time of euthanisation(right). c , f Representative images of immunohistochemistry staining of vimentin, E-cadherin, β-catenin, c-Myc in tumour tissues. g Overall survival HCC patients related to indicated gene expression levels, were generated by the GEPIA2 database via Kaplan–Meier analysis. HCC, hepatocellular carcinoma; TAMs, tumour-associated macrophages; IHC, Immunohistochemistry. Data are presented as mean ± SEM (* p < 0.05, ** p < 0.01 and *** p < 0.001)

Techniques Used: Inhibition, In Vivo, Injection, Immunohistochemistry, Staining, Gene Expression, Generated

Schematic representation illustrates the positive feedback loop between HCC cells and HCC-TAMs. Polarization-promoting factors (IL-10, TGF-β, ect.) in the HCC TME can up-regulate the expression of Wnt2b in macrophages, then promote expression and nuclear translocation of β-catenin, which can promote the M2 polarization of TAMs, a process associated with the activation of HCC-TAMs glycolysis by activating c-Myc. These polarized TAMs can promote the proliferation, migration and EMT of tumor cells. TLR9 agonist CpG ODG can act as a blocker of Wnt2b signal which can inhibit M2 polarization of HCC-TAMs induced by HCC-TCM. HCC, hepatocellular carcinoma; TME, tumor microenvironment; TAMs, tumour-associated macrophages; EMT, epithelial-mesenchymal transformation

Figure Legend Snippet: Schematic representation illustrates the positive feedback loop between HCC cells and HCC-TAMs. Polarization-promoting factors (IL-10, TGF-β, ect.) in the HCC TME can up-regulate the expression of Wnt2b in macrophages, then promote expression and nuclear translocation of β-catenin, which can promote the M2 polarization of TAMs, a process associated with the activation of HCC-TAMs glycolysis by activating c-Myc. These polarized TAMs can promote the proliferation, migration and EMT of tumor cells. TLR9 agonist CpG ODG can act as a blocker of Wnt2b signal which can inhibit M2 polarization of HCC-TAMs induced by HCC-TCM. HCC, hepatocellular carcinoma; TME, tumor microenvironment; TAMs, tumour-associated macrophages; EMT, epithelial-mesenchymal transformation

Techniques Used: Expressing, Translocation Assay, Activation Assay, Migration, Transformation Assay

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Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Promotion of epithelial-mesenchymal transformation by hepatocellular carcinoma-educated macrophages through Wnt2b/β-catenin/c-Myc signaling and reprogramming glycolysis

doi: 10.1186/s13046-020-01808-3

Figure Lengend Snippet: HCC-TCM promotes M2 polarization via Wnt2b/β-catenin signalling. a THP-1 derived macrophages (THP-1-M) were incubated with 50% HCC-TCM for 48 h to obtain the HCC-educated macrophages (HCC-TAMs). Transcript expression levels of Wnt2b were determined in HCC-TAMs by qPCR. b The expression levels of Wnt2b (red) in CD68 + macrophages (green) were determined by immunofluorescence using TMA containing pairs of tumors and matched para-carcinoma tissues of HCC patients. c , d THP-1-M were transfected with control vectors or Wnt2B-V5 (over-Wnt2b) vectors for 48 h. The expression levels of CD163 and markers for M1 or M2 macrophages on/in these cells were determined by flow cytometry and qPCR, respectively. THP-1-M infected with control vectors, sh-Wnt2b or sh-CTNNB1 (β-catenin) vector were acquired as described in the Materials and Methods, and then incubated with 50% HCC-TCM for 48 h. The expression levels of CD163 and markers for M1 or M2 macrophages on/in these cells were determined by flow cytometry ( e , i ) and qPCR ( f , j ), respectively. The expression levels of β-catenin in HCC-TAMs that were infected with control vectors or sh-Wnt2b vectors were determined by western blotting and immunofluorescence respectively ( g , h ). One representative of at least three independent experiments is shown. qPCR, quantitative real-time PCR; HCC, hepatocellular carcinoma; TCM, tumour condition culture medium; TAMs, tumour-associated macrophages; 7721, SMMC-7721; TMA, Tissue microarray. Data are presented as mean ± SEM from at least three independent experiments (* p < 0.05, ** p < 0.01 and *** p < 0.001)

Article Snippet: Active Wnt2B-V5 encoding homo Wnt2b (plasmid #43808; Addgene) was a gift from Xi He (Professor of Neurology, Harvard Medical School, USA).

Techniques: Derivative Assay, Incubation, Expressing, Immunofluorescence, Transfection, Control, Flow Cytometry, Infection, Plasmid Preparation, Western Blot, Real-time Polymerase Chain Reaction, Microarray

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Promotion of epithelial-mesenchymal transformation by hepatocellular carcinoma-educated macrophages through Wnt2b/β-catenin/c-Myc signaling and reprogramming glycolysis

doi: 10.1186/s13046-020-01808-3

Figure Lengend Snippet: The activation of Wnt2b/β-catenin signalling enhances TAMs-induced tumour-promoting effects. a THP-1 derived macrophages (THP-1-M) were transfected with control vectors or Wnt2B-V5 (over-Wnt2b) vectors for 48 h. These macrophages were incubated with RPMI 1640 for an additional 24 h to obtain the condition medium (CM). HCC cells were cultured in the presence of indicated CM for 48 h. The expression levels of EMT markers were determined by western blotting. ( b-e ) THP-1-M infected with control vectors, sh-Wnt2b or sh-CTNNB1 (β-catenin) vectors were acquired as described in Materials and Methods. These macrophages were incubated with 50% HCC-TCM for 48 h for the preparation of the different TAMs. These TAMs were incubated with RPMI 1640 for another 24 h to obtain the CM. b , c HCC cells were cultured in the presence of the indicated CM for 48 h. The expression levels of EMT markers were determined by western blotting. d HCC cells were incubated with culture medium (Ctrl) or the indicated CM for 24 h. The cell viability of each group was detected by MTT assay. e HCC cells were scratched with a plastic pipette tip and incubated with culture medium (Ctrl) or indicated CM for 24 h. The results of this wound healing assay were photographed and measured. One representative of at least three independent experiments is shown. qPCR, quantitative real-time PCR; HCC, hepatocellular carcinoma; TCM, tumour condition culture medium; TAMs, tumour-associated macrophages; 7721, SMMC-7721. Data are presented as mean ± SEM from at least three independent experiments (* p < 0.05, ** p < 0.01 and *** p < 0.001)

Article Snippet: Active Wnt2B-V5 encoding homo Wnt2b (plasmid #43808; Addgene) was a gift from Xi He (Professor of Neurology, Harvard Medical School, USA).

Techniques: Activation Assay, Derivative Assay, Transfection, Control, Incubation, Cell Culture, Expressing, Western Blot, Infection, MTT Assay, Transferring, Wound Healing Assay, Real-time Polymerase Chain Reaction

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Promotion of epithelial-mesenchymal transformation by hepatocellular carcinoma-educated macrophages through Wnt2b/β-catenin/c-Myc signaling and reprogramming glycolysis

doi: 10.1186/s13046-020-01808-3

Figure Lengend Snippet: The inhibition of Wnt2b/β-catenin signaling reduces the tumour-promoting effect of HCC-TAMs in vivo. SMMC-7721 cells (6 × 10 6 ) with or without the indicated TAMs (1.5 × 10 6 ) were mixed with Matrigel (at ratio 4:1) and subcutaneously injected into the right subaxillary of 6-week old immunodeficient mice ( n = 6 mice/group). a , d Representative images of the subcutaneous tumors from each group. b , e Growth of subcutaneous tumours (left); the average tumour weight of each group at the time of euthanisation(right). c , f Representative images of immunohistochemistry staining of vimentin, E-cadherin, β-catenin, c-Myc in tumour tissues. g Overall survival HCC patients related to indicated gene expression levels, were generated by the GEPIA2 database via Kaplan–Meier analysis. HCC, hepatocellular carcinoma; TAMs, tumour-associated macrophages; IHC, Immunohistochemistry. Data are presented as mean ± SEM (* p < 0.05, ** p < 0.01 and *** p < 0.001)

Article Snippet: Active Wnt2B-V5 encoding homo Wnt2b (plasmid #43808; Addgene) was a gift from Xi He (Professor of Neurology, Harvard Medical School, USA).

Techniques: Inhibition, In Vivo, Injection, Immunohistochemistry, Staining, Gene Expression, Generated

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Promotion of epithelial-mesenchymal transformation by hepatocellular carcinoma-educated macrophages through Wnt2b/β-catenin/c-Myc signaling and reprogramming glycolysis

doi: 10.1186/s13046-020-01808-3

Figure Lengend Snippet: Schematic representation illustrates the positive feedback loop between HCC cells and HCC-TAMs. Polarization-promoting factors (IL-10, TGF-β, ect.) in the HCC TME can up-regulate the expression of Wnt2b in macrophages, then promote expression and nuclear translocation of β-catenin, which can promote the M2 polarization of TAMs, a process associated with the activation of HCC-TAMs glycolysis by activating c-Myc. These polarized TAMs can promote the proliferation, migration and EMT of tumor cells. TLR9 agonist CpG ODG can act as a blocker of Wnt2b signal which can inhibit M2 polarization of HCC-TAMs induced by HCC-TCM. HCC, hepatocellular carcinoma; TME, tumor microenvironment; TAMs, tumour-associated macrophages; EMT, epithelial-mesenchymal transformation

Article Snippet: Active Wnt2B-V5 encoding homo Wnt2b (plasmid #43808; Addgene) was a gift from Xi He (Professor of Neurology, Harvard Medical School, USA).

Techniques: Expressing, Translocation Assay, Activation Assay, Migration, Transformation Assay

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