Journal: Cancer Management and Research

Article Title: WAVE2 Enhanced Hepatic Stellate Cells Activity in Colorectal Liver Metastases

doi: 10.2147/CMAR.S259125

Figure Lengend Snippet: Association between WAVE2 protein expression and hepatic micro-metastasis in human CLM. ( A ) Profiles of sinusoid-associated micro-metastasis (SAM) and portal-associated micro-metastasis (PAM) following H&E staining. ( B ) Detection of CLM in hepatic tissue sections via immunohistochemical staining with CK20. ( C ) Profiles of CD31-labeled MVD in CLM. ( D ) Immunohistochemical staining of WAVE2 protein expression in SAM and PAM. Scale bar=50 μm. ( E ) PAM lesions exhibit higher WAVE2 protein levels than SAM lesions. Quantification of WAVE2 expression by H-score is also shown. *** P <0.001. ( F ) The relationship between WAVE2 and MVD in CLM. *** P <0.001.

Article Snippet: Packaging lentiviruses encoding non-targeting short hairpin RNAs (sh-NT) and WAVE2 shRNA (sh-WAVE2) were purchased from Santa Cruz Biotechnology (NM_001201404-Q9Y6W5 and NM_006990-605875; Cat# sc-36833-V), whereas si-YAP was acquired from Guangzhou RiboBio (Guangzhou, China).

Techniques: Expressing, Staining, Immunohistochemical staining, Labeling

Journal: Cancer Management and Research

Article Title: WAVE2 Enhanced Hepatic Stellate Cells Activity in Colorectal Liver Metastases

doi: 10.2147/CMAR.S259125

Figure Lengend Snippet: WAVE2 knockdown inhibits activation of HSCs into tumor-associated myofibroblasts. ( A ) Downregulation of WAVE2 expression via shWAVE2 lentiviral knockdown in human HSCs cells. Western blots showing efficiently-knocked down WAVE2. ( B ) HSCs transduced with shNC or shWAVE2 lentiviruses, treated with TGF-β1 (2.5 ng/mL) and subjected to IF for α-SMA (green). WAVE2 knockdown consistently suppressed TGF-β1 activation of HSCs into myofibroblasts. Bar=50 mm. ** P <0.01, *** P <0.001; n=5 randomly picked microscopic fields. ( C ) Control and WAVE2 knockdown HSCs, serum-starved and treated with TGF-β1 after 24 hours. Cell lysates were subjected to Western blot analysis for detection of HSC activation markers, α-SMA and p-SMAD2.

Article Snippet: Packaging lentiviruses encoding non-targeting short hairpin RNAs (sh-NT) and WAVE2 shRNA (sh-WAVE2) were purchased from Santa Cruz Biotechnology (NM_001201404-Q9Y6W5 and NM_006990-605875; Cat# sc-36833-V), whereas si-YAP was acquired from Guangzhou RiboBio (Guangzhou, China).

Techniques: Knockdown, Activation Assay, Expressing, Western Blot, Transduction, Control

Journal: Cancer Management and Research

Article Title: WAVE2 Enhanced Hepatic Stellate Cells Activity in Colorectal Liver Metastases

doi: 10.2147/CMAR.S259125

Figure Lengend Snippet: WAVE2/YAP1 signaling is a critical driver of activation of HSC processes. ( A ) Western blots of HSC activation showing lower levels of WAVE2, α-SMA and p-SMAD2 in Si-YAP1-transfected HSC cells relative to controls. ( B ) IF analysis for WAVE2 (green) and p-SMAD2 (red).

Article Snippet: Packaging lentiviruses encoding non-targeting short hairpin RNAs (sh-NT) and WAVE2 shRNA (sh-WAVE2) were purchased from Santa Cruz Biotechnology (NM_001201404-Q9Y6W5 and NM_006990-605875; Cat# sc-36833-V), whereas si-YAP was acquired from Guangzhou RiboBio (Guangzhou, China).

Techniques: Activation Assay, Western Blot, Transfection

Journal: Cancer Management and Research

Article Title: WAVE2 Enhanced Hepatic Stellate Cells Activity in Colorectal Liver Metastases

doi: 10.2147/CMAR.S259125

Figure Lengend Snippet: WAVE2 knockdown reduces stimulatory effects of HSCs on tumor prognosis in mice. ( A ) HT29 cells (0.5×10 6 ) mixed with 0.5×10 6 a-HSCs expressing either sh-NC or sh-WAVE2 were implanted into nude mice by orthotropic transplantation. Images of dissected tumors from mice (left) and the tumor size between the HT29+HSC-shNC and HT29+HSC-shWAVE2 groups (right) were compared. Each bar represents mean ± SD of six mice per group. ( B ) IF staining for α-SMA detection showing that WAVE2 knockdown HSCs impairs HT29 tumor growth in mice. *** P <0.001 ( C ) IHC staining of CD31 in xenograft. *** P <0.001.

Article Snippet: Packaging lentiviruses encoding non-targeting short hairpin RNAs (sh-NT) and WAVE2 shRNA (sh-WAVE2) were purchased from Santa Cruz Biotechnology (NM_001201404-Q9Y6W5 and NM_006990-605875; Cat# sc-36833-V), whereas si-YAP was acquired from Guangzhou RiboBio (Guangzhou, China).

Techniques: Knockdown, Expressing, Transplantation Assay, Staining, Immunohistochemistry

Journal: Cancer Management and Research

Article Title: WAVE2 Enhanced Hepatic Stellate Cells Activity in Colorectal Liver Metastases

doi: 10.2147/CMAR.S259125

Figure Lengend Snippet: Schematic diagram showing the effect of WAVE2 on HSC processes and paracrine signaling in the tumor microenvironment. WAVE2 was associated with regulatory functions of the TGF-β1/YAP1 signaling in CLM pathogenesis.

Article Snippet: Packaging lentiviruses encoding non-targeting short hairpin RNAs (sh-NT) and WAVE2 shRNA (sh-WAVE2) were purchased from Santa Cruz Biotechnology (NM_001201404-Q9Y6W5 and NM_006990-605875; Cat# sc-36833-V), whereas si-YAP was acquired from Guangzhou RiboBio (Guangzhou, China).

Techniques:

Journal: Cancer Management and Research

Article Title:

WAVE2 Enhanced Hepatic Stellate Cells Activity in Colorectal Liver Metastases

doi: 10.2147/cmar.s259125

Figure Lengend Snippet: Figure 1 Association between WAVE2 protein expression and hepatic micro-metastasis in human CLM. (A) Profiles of sinusoid-associated micro-metastasis (SAM) and portal-associated micro-metastasis (PAM) following H&E staining. (B) Detection of CLM in hepatic tissue sections via immunohistochemical staining with CK20. (C) Profiles of CD31-labeled MVD in CLM. (D) Immunohistochemical staining of WAVE2 protein expression in SAM and PAM. Scale bar=50 μm. (E) PAM lesions exhibit higher WAVE2 protein levels than SAM lesions. Quantification of WAVE2 expression by H-score is also shown. *** P<0.001. (F) The relationship between WAVE2 and MVD in CLM. *** P<0.001.

Article Snippet: Packaging lentiviruses encoding non-targeting short hairpin RNAs (sh-NT) and WAVE2 shRNA (shWAVE2) were purchased from Santa Cruz Biotechnology (NM_001201404-Q9Y6W5 and NM_006990-605875; Cat# sc-36833-V), whereas si-YAP was acquired from Guangzhou RiboBio (Guangzhou, China).

Techniques: Expressing, Staining, Immunohistochemical staining, Labeling

Journal: Cancer Management and Research

Article Title:

WAVE2 Enhanced Hepatic Stellate Cells Activity in Colorectal Liver Metastases

doi: 10.2147/cmar.s259125

Figure Lengend Snippet: Figure 3 WAVE2/YAP1 signaling is a critical driver of activation of HSC processes. (A) Western blots of HSC activation showing lower levels of WAVE2, α-SMA and p-SMAD2 in Si-YAP1-transfected HSC cells relative to controls. (B) IF analysis for WAVE2 (green) and p-SMAD2 (red).

Article Snippet: Packaging lentiviruses encoding non-targeting short hairpin RNAs (sh-NT) and WAVE2 shRNA (shWAVE2) were purchased from Santa Cruz Biotechnology (NM_001201404-Q9Y6W5 and NM_006990-605875; Cat# sc-36833-V), whereas si-YAP was acquired from Guangzhou RiboBio (Guangzhou, China).

Techniques: Activation Assay, Western Blot, Transfection

Journal: Cancer Management and Research

Article Title:

WAVE2 Enhanced Hepatic Stellate Cells Activity in Colorectal Liver Metastases

doi: 10.2147/cmar.s259125

Figure Lengend Snippet: Figure 2 WAVE2 knockdown inhibits activation of HSCs into tumor-associated myofibroblasts. (A) Downregulation of WAVE2 expression via shWAVE2 lentiviral knockdown in human HSCs cells. Western blots showing efficiently-knocked down WAVE2. (B) HSCs transduced with shNC or shWAVE2 lentiviruses, treated with TGF-β1 (2.5 ng/mL) and subjected to IF for α-SMA (green). WAVE2 knockdown consistently suppressed TGF-β1 activation of HSCs into myofibroblasts. Bar=50 mm. **P<0.01, ***P<0.001; n=5 randomly picked microscopic fields. (C) Control and WAVE2 knockdown HSCs, serum-starved and treated with TGF-β1 after 24 hours. Cell lysates were subjected to Western blot analysis for detection of HSC activation markers, α-SMA and p-SMAD2.

Article Snippet: Packaging lentiviruses encoding non-targeting short hairpin RNAs (sh-NT) and WAVE2 shRNA (shWAVE2) were purchased from Santa Cruz Biotechnology (NM_001201404-Q9Y6W5 and NM_006990-605875; Cat# sc-36833-V), whereas si-YAP was acquired from Guangzhou RiboBio (Guangzhou, China).

Techniques: Knockdown, Activation Assay, Expressing, Western Blot, Transduction, Control

Journal: Cancer Management and Research

Article Title:

WAVE2 Enhanced Hepatic Stellate Cells Activity in Colorectal Liver Metastases

doi: 10.2147/cmar.s259125

Figure Lengend Snippet: Figure 4 WAVE2 knockdown reduces stimulatory effects of HSCs on tumor prognosis in mice. (A) HT29 cells (0.5×106) mixed with 0.5×106 a-HSCs expressing either sh- NC or sh-WAVE2 were implanted into nude mice by orthotropic transplantation. Images of dissected tumors from mice (left) and the tumor size between the HT29+HSC- shNC and HT29+HSC-shWAVE2 groups (right) were compared. Each bar represents mean ± SD of six mice per group. (B) IF staining for α-SMA detection showing that WAVE2 knockdown HSCs impairs HT29 tumor growth in mice. *** P<0.001 (C) IHC staining of CD31 in xenograft. *** P<0.001.

Article Snippet: Packaging lentiviruses encoding non-targeting short hairpin RNAs (sh-NT) and WAVE2 shRNA (shWAVE2) were purchased from Santa Cruz Biotechnology (NM_001201404-Q9Y6W5 and NM_006990-605875; Cat# sc-36833-V), whereas si-YAP was acquired from Guangzhou RiboBio (Guangzhou, China).

Techniques: Knockdown, Expressing, Transplantation Assay, Staining, Immunohistochemistry

Journal: Cancer Management and Research

Article Title:

WAVE2 Enhanced Hepatic Stellate Cells Activity in Colorectal Liver Metastases

doi: 10.2147/cmar.s259125

Figure Lengend Snippet: Figure 5 Schematic diagram showing the effect of WAVE2 on HSC processes and paracrine signaling in the tumor microenvironment. WAVE2 was associated with regulatory functions of the TGF-β1/YAP1 signaling in CLM pathogenesis.

Article Snippet: Packaging lentiviruses encoding non-targeting short hairpin RNAs (sh-NT) and WAVE2 shRNA (shWAVE2) were purchased from Santa Cruz Biotechnology (NM_001201404-Q9Y6W5 and NM_006990-605875; Cat# sc-36833-V), whereas si-YAP was acquired from Guangzhou RiboBio (Guangzhou, China).

Techniques:

Journal: International journal of molecular sciences

Article Title: Gα i 2 Induces Cell Migration in PC3 Prostate Cancer Cells in the Absence of Rac1 Activation.

doi: 10.3390/ijms26062663

Figure Lengend Snippet: Figure 5. (A). Rac1-dependent activation of WAVE2 is impaired in the absence of Gai2. Western blot analysis of Gαi2, p-Wave2 in PC3 cells constitutively overexpressing Rac1, with (+) or without (−) Gαi2 siRNA. Total WAVE2 and α-Tubulin were used as loading controls. (B). Ratios of optical densities (mean ± SEM) of Phosho-Wave2/total Wave2 after different treatments (data from three replicate experiments).

Article Snippet: WAVE2 (Cat# WP1791) were obtained from ECM Biosciences (Versailles, KY, USA).

Techniques: Activation Assay, Western Blot

Journal: Frontiers in Oncology

Article Title: BTG1 Overexpression Might Promote Invasion and Metastasis of Colorectal Cancer via Decreasing Adhesion and Inducing Epithelial–Mesenchymal Transition

doi: 10.3389/fonc.2020.598192

Figure Lengend Snippet: Antibodies used for Western blot.

Article Snippet: WAVE2 (C-6) , Mouse , Santa Cruz Biotechnology.

Techniques: Western Blot

Journal: Heliyon

Article Title: Mangiferin inhibits chronic stress-induced tumor growth in colorectal liver metastases via WAVE2 signaling pathway

doi: 10.1016/j.heliyon.2023.e13753

Figure Lengend Snippet: MGF reduces promoting effects of HSCs on tumor growth in mice. (A) HT-29 cells (0.5 × 10 6 ) mixed with 0.5 × 10 6 a-HSCs expressing tumors were dissected from nude mice, and tumor size was compared between the HT29+HSC-Control and HT29+HSC-MGF groups (left). (B) MVD in HT29+HSC-CUMS and HT29+HSC- MGF group. (C) Masson's trichrome staining of xenograft. (D) IF staining for α-SMA detection shows that MGF inhibits HT-29 tumor cell growth in mice. (E)Western blot showed that expression of β2-AR, WAVE2, VEGF and α-SMA decreased significantly in HT29+HSC-MGF group. Data are presented as the mean ± S.E.M. *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: The stock solution was used to prepare different concentrations in the test media. including β2-AR polyclonal antibody (Cat. #8513, CST), WAVE2 polyclonal antibody (Cat. # YT4898, ImmunoWay), VEGF polyclonal antibody (Cat. # sc-7269, Santa Cruz Biotechnology), α-smooth muscle actin (α-SMA) (Abcam, Cat. # ab5694), a polyclonal antibody against p -SMAD2 (Cat. #3104, CST) at a 1:1000 and GAPDH mouse monoclonal antibody (Cat. # ab8245, Abcam) at 1:5000 dilution.

Techniques: Expressing, Control, Staining, Western Blot

Journal: Heliyon

Article Title: Mangiferin inhibits chronic stress-induced tumor growth in colorectal liver metastases via WAVE2 signaling pathway

doi: 10.1016/j.heliyon.2023.e13753

Figure Lengend Snippet: MGF inhibits activation of HSCs via the WAVE2 signaling pathway. (A) Transduced HSCs treated with TGF-1 (2.5 ng/mL) and IF for SMA show that MGF consistently suppresses TGF-1-induced myofibroblast differentiation. (B) Following 24 h of serum starvation and treatment with TGF-1, control and MGF-treated HSCs are shown. Western blot analysis for detection of HSC activation markers, α-SMA and p -SMAD2, and activator β2-AR and TGF-β1. Data are presented as the mean ± S.E.M. *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: The stock solution was used to prepare different concentrations in the test media. including β2-AR polyclonal antibody (Cat. #8513, CST), WAVE2 polyclonal antibody (Cat. # YT4898, ImmunoWay), VEGF polyclonal antibody (Cat. # sc-7269, Santa Cruz Biotechnology), α-smooth muscle actin (α-SMA) (Abcam, Cat. # ab5694), a polyclonal antibody against p -SMAD2 (Cat. #3104, CST) at a 1:1000 and GAPDH mouse monoclonal antibody (Cat. # ab8245, Abcam) at 1:5000 dilution.

Techniques: Activation Assay, Control, Western Blot