wave2 polyclonal antibody  (ImmunoWay Biotechnology Company)

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

Journal: Cell Cycle

Article Title: SKAP2 regulates Arp2/3 complex for actin-mediated asymmetric cytokinesis by interacting with WAVE2 in mouse oocytes

doi: 10.1080/15384101.2017.1380126

Figure Lengend Snippet: Effects of SKAP2 RNAi on ARP2 and WAVE2 expression. (A) Subcellular localization of ARP2 after SKAP2 siRNA injection. ARP2 was mainly distributed at the membrane in the control oocytes, whereas ARP2 expression was barely detectable in the siRNA-injected group. Green: ARP2; blue: chromatin. Bar = 20 μm. (B) Localization of WAVE2 after SKAP2 siRNA injection. WAVE2 was expressed around the spindle, whereas no specific localization of WAVE2 was observed around spindle in the siRNA-injected group. Red: WAVE2; blue: chromatin. Bar = 20 μm. (C) The fluorescence intensity of ARP2 in the SKAP2 siRNA-injected oocytes was decreased. (D) The fluorescence intensity of WAVE2 in SKAP2 siRNA-injected oocytes was significantly reduced. (E) ARP2 expression was reduced after SKAP2 siRNA injection by western blotting examination, as the relative intensity of ARP2 protein was significantly decreased. (F) WAVE2 expression was decreased after SKAP2 siRNA injection by western blotting analysis, as the relative intensity of WAVE2 protein was significantly reduced. *: significant difference (P < 0.05).

Article Snippet: Rabbit polyclonal anti-WAVE2 antibody (Cat#: 3659) was from Santa Cruz (Santa Cruz, CA, USA).

Techniques: Expressing, Injection, Fluorescence, Western Blot

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

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

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

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

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:

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