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ImmunoWay Biotechnology Company wave2 polyclonal antibody
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, <t>WAVE2,</t> 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.
Wave2 Polyclonal Antibody, supplied by ImmunoWay Biotechnology Company, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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1) Product Images from "Mangiferin inhibits chronic stress-induced tumor growth in colorectal liver metastases via WAVE2 signaling pathway"

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

Journal: Heliyon

doi: 10.1016/j.heliyon.2023.e13753

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.
Figure Legend 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.

Techniques Used: Expressing, Control, Staining, Western Blot

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.
Figure Legend 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.

Techniques Used: Activation Assay, Control, Western Blot



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Santa Cruz Biotechnology rabbit polyclonal anti wave2 antibody
Immunofluorescence analysis was performed to assess the spatial co-localization of SKAP2 and <t>WAVE2</t> during oocyte meiotic maturation. As shown in Fig. 8, SKAP2 exhibited predominant localization in the proximal cortical region of oocytes from metaphase I (MI) to metaphase II (MII) stages. Furthermore, SKAP2-WAVE2 co-localization displayed stage-specific patterns: (1) at the MI stage, concentrated asymmetrically at the cell membrane periphery; (2) during anaphase I (AI), distributed along the emergent cell membrane; and (3) at the MII stage, localized specifically at the intercellular junction between daughter cells. All experiments were repeated at least three times
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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, <t>WAVE2,</t> 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.
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Association between <t>WAVE2</t> 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.
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Association between <t>WAVE2</t> 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.
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Association between <t>WAVE2</t> 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.
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FIGURE 1. Dysbindin-1A and -1C have distinct spatial and temporal expression patterns and dysbindin-1C is not a subunit of the BLOC-1 complex. Tissue extracts from DBA/2J mice were subjected to SDS-PAGE followed by Western blotting using anti-dysbindin-1 antibody. The brain extract from sdy was used as a negative control, and -actin was used as a loading control. These experiments were repeated three times independently. A, dysbindin-1A is widely expressed in multiple mouse tissues, whereas dysbindin-1C is only expressed in the brain and spinal cord. B, in brain sub-regions, the dysbindin-1A levels are higher than dysbindin-1C in the olfactory bulb, substantia nigra, cerebellar cortex, and brain stem, but dysbindin-1C has higher expression levels than dysbindin-1A in the striatum, cerebral cortex, and hippocampal formation. C, dysbindin-1C is mainly enriched in the synaptic vesicles, whereas dysbindin-1A is mainly localized in the presynaptic membrane. In addition, both dysbindin-1A and -1C are found in the proportion of postsynaptic density. Successful synaptic fractionation is confirmed with VAMP2 as a marker for synaptic vesicles and GluR1 as a marker for the postsynaptic density. D and E, protein levels of dysbindin-1A in the hippocampal formation are gradually decreased. In contrast, the dysbindin-1C expression levels increase at postnatal stages. The chart in E is plotted by the relative intensities (IOD) of the bands in D. F, sedimentation velocity analyses. Mouse brain cytosol was fractioned by ultracentrifugation on a 5–20% (w/v) sucrose gradient and probed with antibodies against dysbindin-1, BLOS1, -dystrobrevin, and <t>WAVE2</t> by immunoblotting. Fractions 1 and 20 correspond to the top and bottom ends of the gradient, respectively. Dysbindin-1C does not co-sediment with subunits of the BLOC-1 complex, including dysbindin-1A and BLOS1. Moreover, dysbindin-1C does not form a stable DPC complex with -dystrobrevin nor a stable ternary complex with WAVE2 and Abi-1. Arrowheads, nonspecific bands. G, destabilization of the dysbindin-1 in extracts of three BLOC-1 mutants (sdy, pa, and mu). Sdy is the mutant of dysbindin-1; mu is the mutant of muted; and pa is the mutant of pallidin. Inbred strain DBA/2J served as the control for sdy, CHMU/Le for mu, and C57BL/6J for pa.
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Image Search Results


Immunofluorescence analysis was performed to assess the spatial co-localization of SKAP2 and WAVE2 during oocyte meiotic maturation. As shown in Fig. 8, SKAP2 exhibited predominant localization in the proximal cortical region of oocytes from metaphase I (MI) to metaphase II (MII) stages. Furthermore, SKAP2-WAVE2 co-localization displayed stage-specific patterns: (1) at the MI stage, concentrated asymmetrically at the cell membrane periphery; (2) during anaphase I (AI), distributed along the emergent cell membrane; and (3) at the MII stage, localized specifically at the intercellular junction between daughter cells. All experiments were repeated at least three times

Journal: Reproductive Sciences

Article Title: Expression and Regulatory Roles of SKAP2 and Cortactin in Mouse Ovarian Tissue and Oocyte Maturation

doi: 10.1007/s43032-025-01925-4

Figure Lengend Snippet: Immunofluorescence analysis was performed to assess the spatial co-localization of SKAP2 and WAVE2 during oocyte meiotic maturation. As shown in Fig. 8, SKAP2 exhibited predominant localization in the proximal cortical region of oocytes from metaphase I (MI) to metaphase II (MII) stages. Furthermore, SKAP2-WAVE2 co-localization displayed stage-specific patterns: (1) at the MI stage, concentrated asymmetrically at the cell membrane periphery; (2) during anaphase I (AI), distributed along the emergent cell membrane; and (3) at the MII stage, localized specifically at the intercellular junction between daughter cells. All experiments were repeated at least three times

Article Snippet: Rabbit polyclonal anti-WAVE2 antibody was purchased from Santa Cruz (Santa Cruz, CA, USA).

Techniques: Immunofluorescence, Membrane

To investigate SKAP2-mediated regulation of WAVE2 membrane dynamics, immunofluorescence analysis was performed in control and SKAP2 knockdown oocytes. As shown in Fig. 10, in control groups, WAVE2 exhibited stable expression and cortical localization throughout meiotic progression, with stage-specific co-localization patterns with F-actin: (1) asymmetric cortical clustering at metaphase I (MI), (2) peripheral membrane distribution during anaphase I (AI), and (3) intercellular junction accumulation at metaphase II (MII). However, SKAP2 depletion significantly reduced WAVE2 expression levels and disrupted its co-localization with F-actin across all meiotic stages. All experiments were repeated at least three times. Data are shown as the mean ± standard deviation. The Student’s t-test was used for data analysis. * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001

Journal: Reproductive Sciences

Article Title: Expression and Regulatory Roles of SKAP2 and Cortactin in Mouse Ovarian Tissue and Oocyte Maturation

doi: 10.1007/s43032-025-01925-4

Figure Lengend Snippet: To investigate SKAP2-mediated regulation of WAVE2 membrane dynamics, immunofluorescence analysis was performed in control and SKAP2 knockdown oocytes. As shown in Fig. 10, in control groups, WAVE2 exhibited stable expression and cortical localization throughout meiotic progression, with stage-specific co-localization patterns with F-actin: (1) asymmetric cortical clustering at metaphase I (MI), (2) peripheral membrane distribution during anaphase I (AI), and (3) intercellular junction accumulation at metaphase II (MII). However, SKAP2 depletion significantly reduced WAVE2 expression levels and disrupted its co-localization with F-actin across all meiotic stages. All experiments were repeated at least three times. Data are shown as the mean ± standard deviation. The Student’s t-test was used for data analysis. * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001

Article Snippet: Rabbit polyclonal anti-WAVE2 antibody was purchased from Santa Cruz (Santa Cruz, CA, USA).

Techniques: Membrane, Immunofluorescence, Control, Knockdown, Expressing, Standard Deviation

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.

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

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.

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

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.

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: WAVE2 polyclonal antibodies (Cat. #YT4898, ImmunoWay), including alpha-smooth muscle actin (α-SMA) (abcame, Cat. # ab5831), phosphorylated mothers against decapentaplegic homolog 2 (p-SMAD2) (CST, Cat. # 3101), and transforming growth factor-beta receptor 2 (TβRII) polyclonal antibody (Santa Cruz Biotechnology, Cat. # sc-17791) were diluted at a ratio of 1:1000, whereas β-actin mouse monoclonal antibody (Sigma-Aldrich, Cat. # A1978) was diluted at 1:5000.

Techniques: Expressing, Staining, Immunohistochemical staining, Labeling

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.

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: WAVE2 polyclonal antibodies (Cat. #YT4898, ImmunoWay), including alpha-smooth muscle actin (α-SMA) (abcame, Cat. # ab5831), phosphorylated mothers against decapentaplegic homolog 2 (p-SMAD2) (CST, Cat. # 3101), and transforming growth factor-beta receptor 2 (TβRII) polyclonal antibody (Santa Cruz Biotechnology, Cat. # sc-17791) were diluted at a ratio of 1:1000, whereas β-actin mouse monoclonal antibody (Sigma-Aldrich, Cat. # A1978) was diluted at 1:5000.

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

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).

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: WAVE2 polyclonal antibodies (Cat. #YT4898, ImmunoWay), including alpha-smooth muscle actin (α-SMA) (abcame, Cat. # ab5831), phosphorylated mothers against decapentaplegic homolog 2 (p-SMAD2) (CST, Cat. # 3101), and transforming growth factor-beta receptor 2 (TβRII) polyclonal antibody (Santa Cruz Biotechnology, Cat. # sc-17791) were diluted at a ratio of 1:1000, whereas β-actin mouse monoclonal antibody (Sigma-Aldrich, Cat. # A1978) was diluted at 1:5000.

Techniques: Activation Assay, Western Blot, Transfection

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.

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: WAVE2 polyclonal antibodies (Cat. #YT4898, ImmunoWay), including alpha-smooth muscle actin (α-SMA) (abcame, Cat. # ab5831), phosphorylated mothers against decapentaplegic homolog 2 (p-SMAD2) (CST, Cat. # 3101), and transforming growth factor-beta receptor 2 (TβRII) polyclonal antibody (Santa Cruz Biotechnology, Cat. # sc-17791) were diluted at a ratio of 1:1000, whereas β-actin mouse monoclonal antibody (Sigma-Aldrich, Cat. # A1978) was diluted at 1:5000.

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

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.

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: WAVE2 polyclonal antibodies (Cat. #YT4898, ImmunoWay), including alpha-smooth muscle actin (α-SMA) (abcame, Cat. # ab5831), phosphorylated mothers against decapentaplegic homolog 2 (p-SMAD2) (CST, Cat. # 3101), and transforming growth factor-beta receptor 2 (TβRII) polyclonal antibody (Santa Cruz Biotechnology, Cat. # sc-17791) were diluted at a ratio of 1:1000, whereas β-actin mouse monoclonal antibody (Sigma-Aldrich, Cat. # A1978) was diluted at 1:5000.

Techniques:

FIGURE 1. Dysbindin-1A and -1C have distinct spatial and temporal expression patterns and dysbindin-1C is not a subunit of the BLOC-1 complex. Tissue extracts from DBA/2J mice were subjected to SDS-PAGE followed by Western blotting using anti-dysbindin-1 antibody. The brain extract from sdy was used as a negative control, and -actin was used as a loading control. These experiments were repeated three times independently. A, dysbindin-1A is widely expressed in multiple mouse tissues, whereas dysbindin-1C is only expressed in the brain and spinal cord. B, in brain sub-regions, the dysbindin-1A levels are higher than dysbindin-1C in the olfactory bulb, substantia nigra, cerebellar cortex, and brain stem, but dysbindin-1C has higher expression levels than dysbindin-1A in the striatum, cerebral cortex, and hippocampal formation. C, dysbindin-1C is mainly enriched in the synaptic vesicles, whereas dysbindin-1A is mainly localized in the presynaptic membrane. In addition, both dysbindin-1A and -1C are found in the proportion of postsynaptic density. Successful synaptic fractionation is confirmed with VAMP2 as a marker for synaptic vesicles and GluR1 as a marker for the postsynaptic density. D and E, protein levels of dysbindin-1A in the hippocampal formation are gradually decreased. In contrast, the dysbindin-1C expression levels increase at postnatal stages. The chart in E is plotted by the relative intensities (IOD) of the bands in D. F, sedimentation velocity analyses. Mouse brain cytosol was fractioned by ultracentrifugation on a 5–20% (w/v) sucrose gradient and probed with antibodies against dysbindin-1, BLOS1, -dystrobrevin, and WAVE2 by immunoblotting. Fractions 1 and 20 correspond to the top and bottom ends of the gradient, respectively. Dysbindin-1C does not co-sediment with subunits of the BLOC-1 complex, including dysbindin-1A and BLOS1. Moreover, dysbindin-1C does not form a stable DPC complex with -dystrobrevin nor a stable ternary complex with WAVE2 and Abi-1. Arrowheads, nonspecific bands. G, destabilization of the dysbindin-1 in extracts of three BLOC-1 mutants (sdy, pa, and mu). Sdy is the mutant of dysbindin-1; mu is the mutant of muted; and pa is the mutant of pallidin. Inbred strain DBA/2J served as the control for sdy, CHMU/Le for mu, and C57BL/6J for pa.

Journal: Journal of Biological Chemistry

Article Title: Dysbindin-1C Is Required for the Survival of Hilar Mossy Cells and the Maturation of Adult Newborn Neurons in Dentate Gyrus

doi: 10.1074/jbc.m114.590927

Figure Lengend Snippet: FIGURE 1. Dysbindin-1A and -1C have distinct spatial and temporal expression patterns and dysbindin-1C is not a subunit of the BLOC-1 complex. Tissue extracts from DBA/2J mice were subjected to SDS-PAGE followed by Western blotting using anti-dysbindin-1 antibody. The brain extract from sdy was used as a negative control, and -actin was used as a loading control. These experiments were repeated three times independently. A, dysbindin-1A is widely expressed in multiple mouse tissues, whereas dysbindin-1C is only expressed in the brain and spinal cord. B, in brain sub-regions, the dysbindin-1A levels are higher than dysbindin-1C in the olfactory bulb, substantia nigra, cerebellar cortex, and brain stem, but dysbindin-1C has higher expression levels than dysbindin-1A in the striatum, cerebral cortex, and hippocampal formation. C, dysbindin-1C is mainly enriched in the synaptic vesicles, whereas dysbindin-1A is mainly localized in the presynaptic membrane. In addition, both dysbindin-1A and -1C are found in the proportion of postsynaptic density. Successful synaptic fractionation is confirmed with VAMP2 as a marker for synaptic vesicles and GluR1 as a marker for the postsynaptic density. D and E, protein levels of dysbindin-1A in the hippocampal formation are gradually decreased. In contrast, the dysbindin-1C expression levels increase at postnatal stages. The chart in E is plotted by the relative intensities (IOD) of the bands in D. F, sedimentation velocity analyses. Mouse brain cytosol was fractioned by ultracentrifugation on a 5–20% (w/v) sucrose gradient and probed with antibodies against dysbindin-1, BLOS1, -dystrobrevin, and WAVE2 by immunoblotting. Fractions 1 and 20 correspond to the top and bottom ends of the gradient, respectively. Dysbindin-1C does not co-sediment with subunits of the BLOC-1 complex, including dysbindin-1A and BLOS1. Moreover, dysbindin-1C does not form a stable DPC complex with -dystrobrevin nor a stable ternary complex with WAVE2 and Abi-1. Arrowheads, nonspecific bands. G, destabilization of the dysbindin-1 in extracts of three BLOC-1 mutants (sdy, pa, and mu). Sdy is the mutant of dysbindin-1; mu is the mutant of muted; and pa is the mutant of pallidin. Inbred strain DBA/2J served as the control for sdy, CHMU/Le for mu, and C57BL/6J for pa.

Article Snippet: Other antibodies used in this study were as follows: goat anti-WAVE2 polyclonal antibody (WB, 1:1000, sc-10394, Santa Cruz Biotechnology, Dallas, TX); goat anti- - dystrobrevin polyclonal antibody (WB, 1:200, sc-13815, Santa Cruz Biotechnology); mouse anti- -actin monoclonal antibody (WB, 1:10,000, A5441, Sigma); goat anti-Sox2 polyclonal antibody (IF, 1:1000, sc-17320, Santa Cruz Biotechnology); mouse anti-nestin monoclonal antibody (IF, 1:100, MAB353, Millipore, Billerica, MA); mouse anti-GFAP monoclonal antibody (IF, 1:1000, IF03L, Millipore); mouse anti-GAD67 monoclonal antibody (IF, 1:100, MAB5406, Millipore); mouse anti-calretinin monoclonal antibody (IF, 1:1000, MAB1568, Millipore); rat anti-BrdU monoclonal antibody (IF, 1:100, ab6326, Abcam, Cambridge, UK); goat anti-DCX polyclonal antibody (IF, 1:150, sc-8066, Santa Cruz Biotechnology); mouse anti-NeuN monoclonal antibody (IF, 1:800, MAB377, Millipore); rabbit antiS100 polyclonal antibody (IF, 1:1000, ab868, Abcam); rabbit anti-phospho-CREB (Ser133) polyclonal antibody (IF, 1:200, 9198, Cell Signaling Technology, Danvers, MA), monoclonal mouse anti-Flag antibody (WB, 1:5000, Sigma); and secondary antibody Alexa Fluor 408, 488, or 594 IgG (1:2000, Molecular Probes, Eugene, OR).

Techniques: Expressing, SDS Page, Western Blot, Negative Control, Control, Membrane, Fractionation, Marker, Sedimentation, Mutagenesis