monoclonal mouse anti-human notch1 a6  (BioCarta)

Journal: Oncology Research Featuring Preclinical and Clinical Cancer Therapeutics

Article Title: MicroRNA-744 Inhibits Cellular Proliferation and Invasion of Colorectal Cancer by Directly Targeting Oncogene Notch1

doi: 10.3727/096504018x15188747585738

Figure Lengend Snippet: Figure 3. Notch1 is a direct target of miR-744 in CRC. (A) Predicted and mutated miR-744 binding sequences in the 3¢-untranslated region (3¢-UTR) of Notch1. (B) SW480 and HCT116 cells were cotransfected with miR-744 mimic or miR-NC and p-MIR-WT- Notch1-3¢-UTR or p-MIR-MUT-Notch1-3¢-UTR. At 48 h after transfection, cells were harvested and subjected to the analysis of luciferase activity using the Dual-Luciferase Reporter Assay System. *p < 0.05 versus miR-NC. miR-744 mimics or miR-NC was transfected into SW480 and HCT116 cells. (C) RT-qPCR and (D) Western blot analysis were applied to measure Notch1 mRNA and protein levels, respectively. *p < 0.05 versus miR-NC.

Article Snippet: Subsequent to blocking with 5% nonfat milk in TBS containing 0.1% Tween 20 (TBST), the membranes were incubated overnight at 4°C with mouse anti-human monoclonal Notch1 antibody (1:1,000 dilution; sc-373944; Santa Cruz Biotechnology, Santa Cruz, CA, USA) or mouse anti-human monoclonal b-actin antibody (1:1,000 dilution; sc-69879; Santa Cruz Biotechnology).

Techniques: Binding Assay, Transfection, Luciferase, Activity Assay, Reporter Assay, Quantitative RT-PCR, Western Blot

Journal: Oncology Research Featuring Preclinical and Clinical Cancer Therapeutics

Article Title: MicroRNA-744 Inhibits Cellular Proliferation and Invasion of Colorectal Cancer by Directly Targeting Oncogene Notch1

doi: 10.3727/096504018x15188747585738

Figure Lengend Snippet: Figure 4. Notch1 knockdown reduces the proliferation and invasion of SW480 and HCT116 cells. (A) Notch1 protein expression was measured in SW480 and HCT116 cells transfected with Notch1 small interfering RNA (siRNA) or NC siRNA using Western blot analysis. *p < 0.05 versus NC siRNA. (B) Cell proliferation activity was measured by CCK-8 assay in SW480 and HCT116 cells trans- fected with Notch1 siRNA or NC siRNA. *p < 0.05 versus NC siRNA. (C) Cell invasion capacity was analyzed with Matrigel invasion assay in SW480 and HCT116 cells transfected with Notch1 siRNA or NC siRNA. *p < 0.05 versus NC siRNA.

Article Snippet: Subsequent to blocking with 5% nonfat milk in TBS containing 0.1% Tween 20 (TBST), the membranes were incubated overnight at 4°C with mouse anti-human monoclonal Notch1 antibody (1:1,000 dilution; sc-373944; Santa Cruz Biotechnology, Santa Cruz, CA, USA) or mouse anti-human monoclonal b-actin antibody (1:1,000 dilution; sc-69879; Santa Cruz Biotechnology).

Techniques: Knockdown, Expressing, Transfection, Small Interfering RNA, Western Blot, Activity Assay, CCK-8 Assay, Invasion Assay

Journal: Oncology Research Featuring Preclinical and Clinical Cancer Therapeutics

Article Title: MicroRNA-744 Inhibits Cellular Proliferation and Invasion of Colorectal Cancer by Directly Targeting Oncogene Notch1

doi: 10.3727/096504018x15188747585738

Figure Lengend Snippet: Figure 5. Restoration of Notch1 expression reverses the suppressive effects of miR-744 mimics in CRC cells. miR-744 mimic or miR-NC was introduced into SW480 and HCT116 cells in combination with pcDNA3.1 or pcDNA3.1-Notch1. (A) Western blot analysis was conducted at 72 h posttransfection to determine Notch1 protein level. *p < 0.05 versus miR-NC. #p < 0.05 versus miR-744 mimic + pcDNA3.1-Notch1. (B) CCK-8 and (C) Matrigel invasion assays were used to detect the proliferation and invasion of the above-mentioned treated cells. *p < 0.05 versus miR-NC. #p < 0.05 versus miR-744 mimic + pcDNA3.1-Notch1.

Article Snippet: Subsequent to blocking with 5% nonfat milk in TBS containing 0.1% Tween 20 (TBST), the membranes were incubated overnight at 4°C with mouse anti-human monoclonal Notch1 antibody (1:1,000 dilution; sc-373944; Santa Cruz Biotechnology, Santa Cruz, CA, USA) or mouse anti-human monoclonal b-actin antibody (1:1,000 dilution; sc-69879; Santa Cruz Biotechnology).

Techniques: Expressing, Western Blot, CCK-8 Assay

Journal: Molecular Medicine Reports

Article Title: Special AT-rich sequence-binding protein-1 participates in the maintenance of breast cancer stem cells through regulation of the Notch signaling pathway and expression of Snail1 and Twist1

doi: 10.3892/mmr.2015.3192

Figure Lengend Snippet: Effects of increased or reduced SATB1 expression on Notch1, Notch4, Hes1, Snail1 and Twist1 expression levels in human breast cancer cells. (A) Graph and (B) representative western blot of SATB1, Notch1, Notch4, Hes1, Snail1 and Twist1 expression levels in SATB1-overexpressing and control MCF-7 cells. (C) Graph and (D) representative western blot of SATB1, Notch1, Notch4, Hes1, Snail1 and Twist1 expression levels in SATB1-knockdown and control BT-549 cells. All data are presented as the mean ± standard deviation of experiments in triplicate; * P<0.001 vs. controls. SATB1, special AT-rich sequence-binding protein-1; cont, control; GFP, green fluorescent protein; sh, small hairpin; NC, negative control.

Article Snippet: Blots were incubated in Tris-buffered saline (TBS) (Spectrum Chemical (Shanghai) Co., Ltd) blocking buffer containing 2% milk for 1~2 h at room temperature and then with the mouse monoclonal anti-human antibodies against Notch1 (sc-373891), Hes1 (sc-166410), Snail1 (sc-271977) and Twist1 (sc-81417), as well as rabbit anti-human polyclonal antibodies against SATB1 (sc-28676) and Notch4 (sc-5594) (Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA) diluted 1:200 in TBS with Tween (TBST; containing 0.1% Tween-20 and 2% bovine serum albumin; Spectrum Chemical (Shanghai) Co., Ltd) overnight at 4°C.

Techniques: Expressing, Western Blot, Control, Knockdown, Standard Deviation, Sequencing, Binding Assay, Negative Control

Journal: Molecular Medicine Reports

Article Title: Special AT-rich sequence-binding protein-1 participates in the maintenance of breast cancer stem cells through regulation of the Notch signaling pathway and expression of Snail1 and Twist1

doi: 10.3892/mmr.2015.3192

Figure Lengend Snippet: Effects of SATB1 expression on tumorigenicity and expression levels of Notch1, Notch4, Hes1, Snail1 and Twist1 in vivo. (A) The number of successful tumor engraftments correlated with the number of MCF-7-GFP-cont and MCF-7-SATB1 cells injected during serial dilution. (B) Tumor growth size subsequent to injection of 1×10 6 MCF-7-GFP-cont and MCF-7-SATB1 cells. (C) The number of successful tumor engraftments correlated with the number of BT-549-sh-NC and BT-549-sh-SATB1 cells injected during serial dilution. (D) Tumor growth size subsequent to injection of 1×10 6 BT-549-sh-NC and BT-549-sh-SATB1 cells. Representative immunoblot analysis and expression fold-change of SATB1, Notch1, Notch4, Hes1, Snail1 and Twist1 in (E) MCF-7-GFP-cont and MCF-7-SATB1-derived and (F) BT-549-sh-NC and BT-549-sh-SATB1-derived tumors relative to that of the control group. Representative images of xenografts, hematoxylin and eosin staining and immunohistochemical staining for Ki-67 xenografts, derived from injection of (G) 10 6 MCF-7-GFP-cont and MCF-7-SATB1 cells (magnification, ×10) and (H) 10 6 BT-549-sh-cont and BT-549-sh-SATB1 cells (magnification, ×10). Quantitative data are presented as the mean ± standard deviation for six tumors in each group; * P<0.001 vs. controls. SATB1, special AT-rich sequence-binding protein-1; GFP, green fluorescent protein; cont, control; sh, small hairpin; NC, negative control; relative fold, expression fold-change relative to the control group.

Article Snippet: Blots were incubated in Tris-buffered saline (TBS) (Spectrum Chemical (Shanghai) Co., Ltd) blocking buffer containing 2% milk for 1~2 h at room temperature and then with the mouse monoclonal anti-human antibodies against Notch1 (sc-373891), Hes1 (sc-166410), Snail1 (sc-271977) and Twist1 (sc-81417), as well as rabbit anti-human polyclonal antibodies against SATB1 (sc-28676) and Notch4 (sc-5594) (Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA) diluted 1:200 in TBS with Tween (TBST; containing 0.1% Tween-20 and 2% bovine serum albumin; Spectrum Chemical (Shanghai) Co., Ltd) overnight at 4°C.

Techniques: Expressing, In Vivo, Injection, Serial Dilution, Western Blot, Derivative Assay, Control, Staining, Immunohistochemical staining, Standard Deviation, Sequencing, Binding Assay, Negative Control

Journal: Genes & Cancer

Article Title: Biological significance and therapeutic implication of resveratrol-inhibited Wnt, Notch and STAT3 signaling in cervical cancer cells

doi:

Figure Lengend Snippet: A: Immunocytochemical illustration of intracellular distribution of Notch1, Notch2, Hes1, Wnt2, Wnt5a, β-catenin, p-STAT3 and PIAS3 in HeLa and SiHa cells without (N) and with (R) 100 μM resveratrol treatment for 48h. Immunofluorescent staining results were showed on the up-left corners. B: HE staining in HeLa and SiHa cells without (N) and with (R) 100 μM resveratrol treatment for 48h. C: Trypan blue discrimination of stained (unviable) and unstained (viable) cells. HeLa and SiHa cells were treated with 100 μM resveratrol for 0h, 12h, 24h, 36h and 48h respectively.

Article Snippet: The antibodies used were a mouse anti-human Notch1 monoclonal antibody (1:100, NeoMarkers, Inc, Fremont, California), a rabbit anti-human Notch2 polyclonal antibody (1:100, Santa Cruz, CA, USA), a rabbit anti-human Hes1 polyclonal antibody (1:1000; a generous gift of Tetsuo Sudo, PhD, Toray Industries, Tokyo, Japan), a goat anti-human Wnt2 (1:100; Santa Cruz, CA, USA), a rabbit anti-human Wnt5a polyclonal antibody(1:100; Santa Cruz, CA, USA), a mouse anti-human β-catenin (1:100; Santa Cruz, CA, USA), a rabbit anti-human p-STAT3(1:100; Santa Cruz, CA, USA) and a rabbit anti-human PIAS3 polyclonal antibody (1:100; Santa Cruz, CA, USA).

Techniques: Staining

Journal: Genes & Cancer

Article Title: Biological significance and therapeutic implication of resveratrol-inhibited Wnt, Notch and STAT3 signaling in cervical cancer cells

doi:

Figure Lengend Snippet: A: Evaluation of Notch1, Notch2, Hes1, Wnt2, Wnt5a, β-catenin, p-STAT3 (STAT3) and PIAS3 in HeLa and SiHa cells without (N) and with (R) 100 μM resveratrol treatment for 48h by Western blotting and B: RT-PCR, β-actin was used as a quantitative control. Gray analyses of the Western blotting and RT-PCR results were under each picture, the ordinate was the ratio of each index and β-actin grey value.

Article Snippet: The antibodies used were a mouse anti-human Notch1 monoclonal antibody (1:100, NeoMarkers, Inc, Fremont, California), a rabbit anti-human Notch2 polyclonal antibody (1:100, Santa Cruz, CA, USA), a rabbit anti-human Hes1 polyclonal antibody (1:1000; a generous gift of Tetsuo Sudo, PhD, Toray Industries, Tokyo, Japan), a goat anti-human Wnt2 (1:100; Santa Cruz, CA, USA), a rabbit anti-human Wnt5a polyclonal antibody(1:100; Santa Cruz, CA, USA), a mouse anti-human β-catenin (1:100; Santa Cruz, CA, USA), a rabbit anti-human p-STAT3(1:100; Santa Cruz, CA, USA) and a rabbit anti-human PIAS3 polyclonal antibody (1:100; Santa Cruz, CA, USA).

Techniques: Western Blot, Reverse Transcription Polymerase Chain Reaction

Journal: Genes & Cancer

Article Title: Biological significance and therapeutic implication of resveratrol-inhibited Wnt, Notch and STAT3 signaling in cervical cancer cells

doi:

Figure Lengend Snippet: Immunohistochemical profiling of Notch1, Notch2, Hes1, Wnt2, Wnt5a, β-catenin, p-STAT3 and PIAS3 expression in human cervical tissues (N: normal cervical tissues removed from the uterine fibroidpatients at post-reproduction ages, AC: cervical adenocarcinomas, SC: cervical squamous cell carcinomas. * p <0.01)

Article Snippet: The antibodies used were a mouse anti-human Notch1 monoclonal antibody (1:100, NeoMarkers, Inc, Fremont, California), a rabbit anti-human Notch2 polyclonal antibody (1:100, Santa Cruz, CA, USA), a rabbit anti-human Hes1 polyclonal antibody (1:1000; a generous gift of Tetsuo Sudo, PhD, Toray Industries, Tokyo, Japan), a goat anti-human Wnt2 (1:100; Santa Cruz, CA, USA), a rabbit anti-human Wnt5a polyclonal antibody(1:100; Santa Cruz, CA, USA), a mouse anti-human β-catenin (1:100; Santa Cruz, CA, USA), a rabbit anti-human p-STAT3(1:100; Santa Cruz, CA, USA) and a rabbit anti-human PIAS3 polyclonal antibody (1:100; Santa Cruz, CA, USA).

Techniques: Immunohistochemical staining, Expressing

Journal: Genes & Cancer

Article Title: Biological significance and therapeutic implication of resveratrol-inhibited Wnt, Notch and STAT3 signaling in cervical cancer cells

doi:

Figure Lengend Snippet: A: Tissue microarray-based immunohistochemical staining for Notch1, Notch2, Hes1, Wnt2, Wnt5a, β-catenin, p-STAT3 and PIAS3 in normal cervical tissues removed from the uterine fibroid patients at post-reproduction ages (N), cervical adenocarcinomas (AC) and cervical squamous cell carcinomas (SC) (original magnifications×400). B: Histogram of Table , *p<0.01.

Article Snippet: The antibodies used were a mouse anti-human Notch1 monoclonal antibody (1:100, NeoMarkers, Inc, Fremont, California), a rabbit anti-human Notch2 polyclonal antibody (1:100, Santa Cruz, CA, USA), a rabbit anti-human Hes1 polyclonal antibody (1:1000; a generous gift of Tetsuo Sudo, PhD, Toray Industries, Tokyo, Japan), a goat anti-human Wnt2 (1:100; Santa Cruz, CA, USA), a rabbit anti-human Wnt5a polyclonal antibody(1:100; Santa Cruz, CA, USA), a mouse anti-human β-catenin (1:100; Santa Cruz, CA, USA), a rabbit anti-human p-STAT3(1:100; Santa Cruz, CA, USA) and a rabbit anti-human PIAS3 polyclonal antibody (1:100; Santa Cruz, CA, USA).

Techniques: Microarray, Immunohistochemical staining, Staining

Journal: Genes & Cancer

Article Title: Biological significance and therapeutic implication of resveratrol-inhibited Wnt, Notch and STAT3 signaling in cervical cancer cells

doi:

Figure Lengend Snippet: The primer sequences of polymerase chain reactions

Article Snippet: The antibodies used were a mouse anti-human Notch1 monoclonal antibody (1:100, NeoMarkers, Inc, Fremont, California), a rabbit anti-human Notch2 polyclonal antibody (1:100, Santa Cruz, CA, USA), a rabbit anti-human Hes1 polyclonal antibody (1:1000; a generous gift of Tetsuo Sudo, PhD, Toray Industries, Tokyo, Japan), a goat anti-human Wnt2 (1:100; Santa Cruz, CA, USA), a rabbit anti-human Wnt5a polyclonal antibody(1:100; Santa Cruz, CA, USA), a mouse anti-human β-catenin (1:100; Santa Cruz, CA, USA), a rabbit anti-human p-STAT3(1:100; Santa Cruz, CA, USA) and a rabbit anti-human PIAS3 polyclonal antibody (1:100; Santa Cruz, CA, USA).

Techniques: Sequencing, Countercurrent Chromatography

Journal: The Journal of Neuroscience

Article Title: Increased Generation of Neuronal Progenitors after Ischemic Injury in the Aged Adult Human Forebrain

doi: 10.1523/JNEUROSCI.4667-06.2006

Figure Lengend Snippet: Cellular architecture and proliferative capacity of the adult human SVZ. a, Schematic overview of a human brain hemisphere with the dotted red line indicating the area of analysis. b, c, Representative histology of the SVZ with a gap between the ependymal layer and astrocytic ribbon in the medial part (b) and with the astrocytic ribbon being continuous from the ependymal layer to the brain parenchyma in the dorsal and ventral areas (c). d, Occasionally, PSA-NCAM-positive cells can be detected close to the ependyma or in the astrocytic ribbon and beyond. e, f, Molecules that were shown to regulate neurogenesis such as notch1 (e) and Flk-1/VEGFR-2 (f) are expressed in the ependymal cell layer. g, Proliferating Ki-67-positive cells are often closely associated with the ependymal layer. h, i, Lack of costaining with pan-hematopoietic markers such as CD45 (h) rules out a non-neural origin of the proliferating Ki-67-positive cells (i). Scale bars: b, c, 50 μm; d, g, h, 10 μm; e, f, 25 μm; i, 20 μm.

Article Snippet: The following antibodies were used: mouse anti-β-III-tubulin (clone Tuj1 1:500; Covance), monoclonal mouse anti-polysialic acid neural cell adhesion molecule (PSA-NCAM, clone 2–2B, 1:2000; Millipore, Temecula, CA), monoclonal mouse anti-human notch1 (1:2000, Ab-1, clone A6; Biocarta Europe, Hamburg, Germany), polyclonal rabbit anti-vascular endothelial growth factor -receptor-2 (VEGFR2)/fetal liver kinase receptor 1 (Flk-1) (1:100; Zytomed, Berlin, Germany), monoclonal mouse anti-glial fibrillary acidic protein (GFAP, 1:500; Millipore), polyclonal rabbit anti-GFAP (1:1000; Dako, Carpinteria, CA), monoclonal mouse anti-human CD45/leukocyte common antigen (1:50, clones 2B11+PD7/26; Dako), monoclonal mouse anti-human CD68 (clone PG-M1, 1:100; Dako), monoclonal mouse anti-human CD34 (clone QBEND10, 1:100; Immunotech, Marseille, France), polyclonal rabbit anti-human S100β (1:100; Dako), monoclonal mouse anti-human nestin (1:200; Millipore), polyclonal rabbit anti-Mash1 (1:100; Millipore), polyclonal rabbit anti-Musashi (1:100; Millipore), and monoclonal mouse anti-human Ki-67 (clone MIB-1, 1:50; Dako).

Techniques: