primary antibodies against notch1  (Proteintech)

Journal: Biomedical Reports

Article Title: Notch signaling pathway regulates the progression of fetal growth restriction through mediating immune dysfunction

doi: 10.3892/br.2025.1989

Figure Lengend Snippet: The expression and distribution of Notch1 and Jagged1 is decreased in PBMCs and placenta. (A-E) The expression of Notch1 and Jagged1 in PBMCs and placenta was detected by (A and B) reverse transcription-quantitative PCR, (C) western blotting, (D) immunofluorescence and (E) immunohistochemistry, respectively. ** P<0.01 and *** P<0.001 vs. control. Scale bar, 100 µm. PBMCs, peripheral blood mononuclear cells; FGR, fetal growth restriction.

Article Snippet: After antigen retrieval, the samples were incubated overnight at 4 ̊C with primary antibodies against Notch1 (1:200), Jagged1 (1:200), CD3 (1:200; cat. no. 17617-1-AP), CD86 (1:200; cat. no. 26903-1-AP), CD206 (1:200; cat. no. 18704-1-AP; last 3 obtained from Proteintech Group, Inc.) and Forkhead Box protein 3 (Foxp3; 1:200; cat. no. ab36607; Abcam).

Techniques: Expressing, Reverse Transcription, Real-time Polymerase Chain Reaction, Western Blot, Immunofluorescence, Immunohistochemistry, Control

Journal: Diagnostics

Article Title: NOTCH1- and CD117-Positive Stem Cells in Human Endometriosis and Adenomyosis Lesions

doi: 10.3390/diagnostics14151642

Figure Lengend Snippet: HE and IHC staining of a representative intestinal endometriosis lesion. ( A ) Intestinal endometriosis (within the ileum) with presence of cystic ectopic endometrial glands within the tunica muscularis propria; ( B ) intestinal endometriosis (purple) affecting the intestinal mucosa (left); ( C ) estrogen receptor expression in ectopic endometrial glands; ( D ) CD10 immunopositivity demarcating ectopic endometrial stroma; ( E ) NOTCH1+ cells (dark brown) in ectopic endometrial stroma (bottom) and intracystic debris; ( F ) CD117+ cells in ectopic endometrial stroma surrounding the ectopic cystic formation.

Article Snippet: The next step was incubation with 0.4% casein in phosphate-buffered saline (PBS) to reduce non-specific binding of primary antibody, followed by 1 h incubation at 37 °C with rabbit polyclonal primary antibodies against NOTCH1 (E-AB-12815, Elabscience, Houston, TX, USA), CD117 (RB-9038-RQ, Epredia, Portsmouth, NH, USA), estrogen receptor (1-ES006-07, Quartett, Berlin, Germany) and mouse monoclonal antibody against CD10 (DAK-CD10 IR 786, Dako, Nowy Sącz, Poland).

Techniques: Immunohistochemistry, Expressing

Journal: Diagnostics

Article Title: NOTCH1- and CD117-Positive Stem Cells in Human Endometriosis and Adenomyosis Lesions

doi: 10.3390/diagnostics14151642

Figure Lengend Snippet: HE and IHC staining of an adenomyosis lesion with phasic changes. ( A ) Adenomyosis lesion with secretory changes* in the glands (apical and basal vacuoles); ( B ) CD10 marker visualizing endometrial stromal cells (dark brown stain) in the ectopic lesion; ( C ) NOTCH1+ cells within the ectopic endometrial stroma; ( D ) CD117+ cells within the ectopic endometrial stroma. * Phasic changes are a rare feature of adenomyosis. In such cases, it might be preferable to refer to the lesion as myometrial endometriosis.

Article Snippet: The next step was incubation with 0.4% casein in phosphate-buffered saline (PBS) to reduce non-specific binding of primary antibody, followed by 1 h incubation at 37 °C with rabbit polyclonal primary antibodies against NOTCH1 (E-AB-12815, Elabscience, Houston, TX, USA), CD117 (RB-9038-RQ, Epredia, Portsmouth, NH, USA), estrogen receptor (1-ES006-07, Quartett, Berlin, Germany) and mouse monoclonal antibody against CD10 (DAK-CD10 IR 786, Dako, Nowy Sącz, Poland).

Techniques: Immunohistochemistry, Marker, Staining

Journal: Diagnostics

Article Title: NOTCH1- and CD117-Positive Stem Cells in Human Endometriosis and Adenomyosis Lesions

doi: 10.3390/diagnostics14151642

Figure Lengend Snippet: Box plots representing the percentage of studied stem cells in the stroma of patients with endometriosis and adenomyosis. ( A ) NOTCH1-positive cells in endometriosis ( n = 23) and adenomyosis lesions ( n = 20); ( B ) CD117-positive endometrial cells in endometriosis ( n = 23) and adenomyosis lesions ( n = 20).

Article Snippet: The next step was incubation with 0.4% casein in phosphate-buffered saline (PBS) to reduce non-specific binding of primary antibody, followed by 1 h incubation at 37 °C with rabbit polyclonal primary antibodies against NOTCH1 (E-AB-12815, Elabscience, Houston, TX, USA), CD117 (RB-9038-RQ, Epredia, Portsmouth, NH, USA), estrogen receptor (1-ES006-07, Quartett, Berlin, Germany) and mouse monoclonal antibody against CD10 (DAK-CD10 IR 786, Dako, Nowy Sącz, Poland).

Techniques:

Journal: Diagnostics

Article Title: NOTCH1- and CD117-Positive Stem Cells in Human Endometriosis and Adenomyosis Lesions

doi: 10.3390/diagnostics14151642

Figure Lengend Snippet: IHC showing NOTCH1 and CD117 expression in representative endometriosis and adenomyosis lesions. ( A ) NOTCH1-positive stromal cells (brown) in a representative tubal endometriosis lesion; ( B ) CD117-positive stromal cells (brown) in the same tubal endometriosis lesion; ( C ) spatial plot of the two types of stem cells in the same endometriotic lesion (NOTCH1+ cells—red, CD117+ cells—blue, ectopic endometrial stromal cells—green, lines—nearest neighbor between the two stem cell types); ( D ) NOTCH1-positive stromal cells (brown) in representative adenomyosis lesion; ( E ) CD117-positive stromal cells (brown) in the same adenomyosis lesion; ( F ) spatial plot of the two types of stem cells in the same adenomyosis lesion (NOTCH1+ cells—red, CD117+ cells—blue, ectopic endometrial stromal cells—green, lines—nearest neighbor between the two stem cell types).

Article Snippet: The next step was incubation with 0.4% casein in phosphate-buffered saline (PBS) to reduce non-specific binding of primary antibody, followed by 1 h incubation at 37 °C with rabbit polyclonal primary antibodies against NOTCH1 (E-AB-12815, Elabscience, Houston, TX, USA), CD117 (RB-9038-RQ, Epredia, Portsmouth, NH, USA), estrogen receptor (1-ES006-07, Quartett, Berlin, Germany) and mouse monoclonal antibody against CD10 (DAK-CD10 IR 786, Dako, Nowy Sącz, Poland).

Techniques: Expressing

Journal: Brain Sciences

Article Title: Targeting Microglia/Macrophages Notch1 Protects Neurons from Pyroptosis in Ischemic Stroke

doi: 10.3390/brainsci13121657

Figure Lengend Snippet: Primers for RT–PCR.

Article Snippet: Finally, they were incubated overnight at 4 °C with primary antibodies against CD68 (diluted 1:200, 14-0688-82, Invitrogen, Waltham, MA, USA), Notch1 (diluted 1:200, MA5-32080, Invitrogen), Ym1 (diluted 1:200, 60130, STEMCELL Technologies, Vancouver, BC, Canada), iNOS (diluted 1:200, GB11119-100, Servicebio, Wuhan, China) and Iba1 (diluted 1:500, GB12105, Servicebio, Wuhan, China).

Techniques:

Journal: Brain Sciences

Article Title: Targeting Microglia/Macrophages Notch1 Protects Neurons from Pyroptosis in Ischemic Stroke

doi: 10.3390/brainsci13121657

Figure Lengend Snippet: Notch1 signaling is closely associated with ischemia–reperfusion injury in the mouse brain. ( A ) Representative Western blot images of Notch1, NLRP3, IL1β, and Cl-caspase1 expression in the ischemic penumbra of wild-type mice 3 days after MCAO or sham surgery (n = 5 mice/group). ( B ) Quantification of the relative protein expression of Notch1, NLRP3, IL1β, and Cl-caspase1 in the ischemic penumbra in the MCAO group compared with the sham-operated group. ( C ) The effects of MCAO and sham operation on Notch1 and NLRP3 mRNA levels in the ischemic penumbra of wild-type mice 3 days after the operation. ( D ) Representative images of immunofluorescence staining with antibodies against Notch1 (green) and Iba1 (red). Cell nuclei were labeled with DAPI (blue). n = 5 mice/group. ( E ) The number of Iba1 + Notch1 + cells. ( F ) Representative Western blot images Notch1 and NLRP3 expression in BV2 cells subjected to OGD/R and control group. ( G ) Quantification of the relative protein expression of Notch1 and NLRP3 in BV2 cells subjected to OGD/R and control group. ( H ) Representative Western blot images Notch1, NLRP3, IL1β and Cl-caspase1 expression in HT22 cells subjected to OGD/R and control group. ( I ) Quantification of the relative protein expression of Notch1, NLRP3, IL1β and Cl-caspase1 in HT22 cells subjected to OGD/R and control group. ( J ) mRNA levels of Notch1 and NLRP3 in HT22 cells subjected to OGD/R and control HT22 cells. Mean ± SD. n = 5 in each group. ** p < 0.01, *** p < 0.001. Scale bar, 20 μm.

Article Snippet: Finally, they were incubated overnight at 4 °C with primary antibodies against CD68 (diluted 1:200, 14-0688-82, Invitrogen, Waltham, MA, USA), Notch1 (diluted 1:200, MA5-32080, Invitrogen), Ym1 (diluted 1:200, 60130, STEMCELL Technologies, Vancouver, BC, Canada), iNOS (diluted 1:200, GB11119-100, Servicebio, Wuhan, China) and Iba1 (diluted 1:500, GB12105, Servicebio, Wuhan, China).

Techniques: Western Blot, Expressing, Immunofluorescence, Staining, Labeling

Journal: Brain Sciences

Article Title: Targeting Microglia/Macrophages Notch1 Protects Neurons from Pyroptosis in Ischemic Stroke

doi: 10.3390/brainsci13121657

Figure Lengend Snippet: Notch1 knockout attenuates CIRI and cerebral migration of monocyte-derived macrophages. ( A ) TTC staining for cerebral infarct analysis. ( B ) Statistical analysis of the infarct volume according to TTC staining. The infarct size is expressed as percentage of infarct volume to contralateral hemisphere size 3 days after MCAO (n = 6 mice/group). ( C ) Statistical analysis of neurological function scores 3 days after MCAO (n = 6 mice/group). ( D ) Statistical analysis of the number of Iba1 + cells. ( E ) Representative images of immunofluorescence staining with an antibody against Iba1 (red). Cell nuclei were labeled with DAPI (blue). (n = 5 mice/group) ( F ) Gating strategy to identify CD45 inter CD11b + cells (MiDM) and CD45 high CD11b + cells (MDMs). ( G ) Statistical analyses of the numbers of MiDM, MDMs and total monocytes. (n = 5 mice/group) Mean ± SD. ** p < 0.01, *** p < 0.001. Scale bar, 20 μm.

Article Snippet: Finally, they were incubated overnight at 4 °C with primary antibodies against CD68 (diluted 1:200, 14-0688-82, Invitrogen, Waltham, MA, USA), Notch1 (diluted 1:200, MA5-32080, Invitrogen), Ym1 (diluted 1:200, 60130, STEMCELL Technologies, Vancouver, BC, Canada), iNOS (diluted 1:200, GB11119-100, Servicebio, Wuhan, China) and Iba1 (diluted 1:500, GB12105, Servicebio, Wuhan, China).

Techniques: Knock-Out, Migration, Derivative Assay, Staining, Immunofluorescence, Labeling

Journal: Brain Sciences

Article Title: Targeting Microglia/Macrophages Notch1 Protects Neurons from Pyroptosis in Ischemic Stroke

doi: 10.3390/brainsci13121657

Figure Lengend Snippet: Notch1 knockout promotes M2 phenotypes in the ischemic penumbra. ( A ) Representative images of immunofluorescence staining with antibodies against CD68 (green) and iNOS (red). ( B ) Statistical analysis of the number of CD68 + iNOS + cells. ( C ) Representative images of immunofluorescence staining with antibodies against CD68 (green) and YM1 (red). Cell nuclei were labeled with DAPI (blue). ( D ) Statistical analysis of the number of CD68 + YM1 + cells. ( E ) Representative Western blot bands and ( F ) quantitative analysis; target protein levels were normalized to β-actin levels. Mean ± SD. n = 5 in each group. * p < 0.05, ** p < 0.01, *** p < 0.001. Scale bar, 20 μm.

Article Snippet: Finally, they were incubated overnight at 4 °C with primary antibodies against CD68 (diluted 1:200, 14-0688-82, Invitrogen, Waltham, MA, USA), Notch1 (diluted 1:200, MA5-32080, Invitrogen), Ym1 (diluted 1:200, 60130, STEMCELL Technologies, Vancouver, BC, Canada), iNOS (diluted 1:200, GB11119-100, Servicebio, Wuhan, China) and Iba1 (diluted 1:500, GB12105, Servicebio, Wuhan, China).

Techniques: Knock-Out, Immunofluorescence, Staining, Labeling, Western Blot

Journal: Brain Sciences

Article Title: Targeting Microglia/Macrophages Notch1 Protects Neurons from Pyroptosis in Ischemic Stroke

doi: 10.3390/brainsci13121657

Figure Lengend Snippet: Notch1 knockout alleviates CIRI-induced pyroptosis. ( A ) Representative Western blot bands and ( B ) Quantitative analysis; target protein levels were normalized to β-actin levels. ( C ) mRNA levels of inflammasome components (NLRP3) and pyroptosis-related proteins (IL-1β and IL-18) in Notch1 FL/FL and Notch1 M-KO mice in the MCAO group and sham group. Mean ± SD. n = 5 in each group. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Finally, they were incubated overnight at 4 °C with primary antibodies against CD68 (diluted 1:200, 14-0688-82, Invitrogen, Waltham, MA, USA), Notch1 (diluted 1:200, MA5-32080, Invitrogen), Ym1 (diluted 1:200, 60130, STEMCELL Technologies, Vancouver, BC, Canada), iNOS (diluted 1:200, GB11119-100, Servicebio, Wuhan, China) and Iba1 (diluted 1:500, GB12105, Servicebio, Wuhan, China).

Techniques: Knock-Out, Western Blot

Journal: Brain Sciences

Article Title: Targeting Microglia/Macrophages Notch1 Protects Neurons from Pyroptosis in Ischemic Stroke

doi: 10.3390/brainsci13121657

Figure Lengend Snippet: BV2 microglia regulate OGD/R-induced apoptosis in HT22 cells via Notch1 in vitro. ( A ) Schematic diagram of the BV2 cell and HT22 cell co-culture system. BV2 cells stably transfected with NC-siRNA lentivirus or Notch1-siRNA lentivirus were plated in the upper chamber, and HT22 hippocampal neurons were plated in the lower chamber. After 24 h, the co-cultured cells were subjected to OGD/R, and HT22 cells in the lower chamber were collected 12 h after reoxygenation for analysis. ( B ) Flow cytometry analysis of apoptosis. ( C ) Statistical analysis of the percentage of apoptotic cells. ( D ) Analysis of relative cell viability by the CCK-8 assay. ( E ) Western blot analysis of Bcl2 and Bax protein expression. ( F ) Relative quantification of Bcl2 and Bax protein expression. Mean ± SD. n = 5 in each group. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Finally, they were incubated overnight at 4 °C with primary antibodies against CD68 (diluted 1:200, 14-0688-82, Invitrogen, Waltham, MA, USA), Notch1 (diluted 1:200, MA5-32080, Invitrogen), Ym1 (diluted 1:200, 60130, STEMCELL Technologies, Vancouver, BC, Canada), iNOS (diluted 1:200, GB11119-100, Servicebio, Wuhan, China) and Iba1 (diluted 1:500, GB12105, Servicebio, Wuhan, China).

Techniques: In Vitro, Co-Culture Assay, Stable Transfection, Transfection, Cell Culture, Flow Cytometry, CCK-8 Assay, Western Blot, Expressing

Journal: Brain Sciences

Article Title: Targeting Microglia/Macrophages Notch1 Protects Neurons from Pyroptosis in Ischemic Stroke

doi: 10.3390/brainsci13121657

Figure Lengend Snippet: Notch1 knockdown in microglial BV2 cells inhibits OGD/R-induced JAK2/STAT3 activation and pyroptosis in HT22 cells. ( A ) Western blot analysis of the expression of pyroptosis-associated proteins in HT22 cells co-cultured with NC-siRNA-transfected BV2 cells and HT22 cells co-cultured with Notch1-siRNA-transfected BV2 cells. ( B ) Relative quantification of pyroptosis-associated protein expression in HT22 cells. ( C ) mRNA expression levels of inflammasome-associated proteins (NLRP3 and Cl-caspase1) and pyroptosis-associated proteins (Cl-GSDMD, IL-1β and IL-18) in HT22 cells co-cultured with NC-siRNA-transfected BV2 cells and those co-cultured with Notch1-siRNA-transfected BV2 cells after OGD/R or sham treatment. ( D ) Protein–protein interaction (PPI) network of Notch1 and JAK2/STAT3. ( E ) Representative Western blot images of P-JAK2, JAK2, P-STAT3, and STAT3 in HT22 cells co-cultured with NC-siRNA-transfected BV2 cells and HT22 cells co-cultured with Notch1-siRNA-transfected BV2 cells after OGD/R or sham treatment. ( F ) Relative quantification of P-JAK2 and P-STAT3 protein expression. Mean ± SD. n = 5 in each group. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Finally, they were incubated overnight at 4 °C with primary antibodies against CD68 (diluted 1:200, 14-0688-82, Invitrogen, Waltham, MA, USA), Notch1 (diluted 1:200, MA5-32080, Invitrogen), Ym1 (diluted 1:200, 60130, STEMCELL Technologies, Vancouver, BC, Canada), iNOS (diluted 1:200, GB11119-100, Servicebio, Wuhan, China) and Iba1 (diluted 1:500, GB12105, Servicebio, Wuhan, China).

Techniques: Activation Assay, Western Blot, Expressing, Cell Culture, Transfection

Journal: Frontiers in Immunology

Article Title: Sulindac sulfide as a non-immune suppressive γ-secretase modulator to target triple-negative breast cancer

doi: 10.3389/fimmu.2023.1244159

Figure Lengend Snippet: SS has gamma secretase modulator (GSM) activity and inhibits Notch1 cleavage. APP C99 and NOTCH1 were transiently transfected into HEK 293T wild type, PS1, and PS2 KO cells. After 16 hours, cells were treated with 5, 12.5, 25, 50, and 75 μM SS or SF in fresh media. Conditioned media were collected after 24 hours and assayed by Aβ ELISA as described in the method section (A, B) . Similarly, H4 cells stably overexpress Notch1 or APP C99 were treated with 25, 50, 75, and 100 μM SS; conditioned media were collected after 24 hours and assayed by Aβ ELISA (C) . Human MDA-MB-231 cells were treated with 10, 25, 50, and 100 μM SS for 48 hours, after which the expression of total Notch1 and cleaved Notch1 (CN1) was measured by Western Blot analysis (D) .

Article Snippet: Membranes were incubated with primary antibodies against Notch1 (D1E11), Notch1-IC (Val1744:D3B8) (Cell Signaling), GAPDH, and β-tubulin (Santa Cruz Biotechnology).

Techniques: Activity Assay, Transfection, Enzyme-linked Immunosorbent Assay, Stable Transfection, Expressing, Western Blot

Journal: Frontiers in Immunology

Article Title: Sulindac sulfide as a non-immune suppressive γ-secretase modulator to target triple-negative breast cancer

doi: 10.3389/fimmu.2023.1244159

Figure Lengend Snippet: SS-mediated anti-mammosphere activity depends on Notch expression. Vector control and intracellular Notch1-overexpressing (N1IC) MDA-MB-231 cells (10,000) were grown in Mammocult media and treated with SS (10 or 50 μM) for one week (twice/week). Following incubation, mammospheres were counted using a Nikon microscope. Representative photographs and average mammospheres sizes (areas) are presented in (A, B) respectively. Data are means ± SD; P-values: *** P < 0.001; **** P < 0.0001, student t-test, using GraphPad Prism. ns, not significant.

Article Snippet: Membranes were incubated with primary antibodies against Notch1 (D1E11), Notch1-IC (Val1744:D3B8) (Cell Signaling), GAPDH, and β-tubulin (Santa Cruz Biotechnology).

Techniques: Activity Assay, Expressing, Plasmid Preparation, Control, Incubation, Microscopy

Journal: Frontiers in Immunology

Article Title: Sulindac sulfide as a non-immune suppressive γ-secretase modulator to target triple-negative breast cancer

doi: 10.3389/fimmu.2023.1244159

Figure Lengend Snippet: SS monotherapy inhibits the growth of a syngeneic TNBC model (C0321). Mouse TNBC C0321 cells (1 million) were injected into the mammary fat pads of syngeneic immunocompetent FVB (female) mice with 1:1 ratio of Matrigel. Palpable tumors were treated with vehicle or SS (60mg/kg, daily, PO) for another two weeks. Tumor volumes and weights were measured twice per week, and three weeks after tumor inoculation, tumors were harvested, weighed, and analyzed by H&E and immunohistochemistry for Notch1 and Jagged1 (A, B) . Fresh tumor specimens were dissociated by Liberase digestion, and single-cell suspensions were analyzed for tumor-infiltrating T-Cells (CD4 and CD8), Dendritic cells (CD11c), MDSC, and TAM (C) by Flow cytometer. All cells were gated on pan-leukocyte marker CD45. Data are means ± SD; P-values: * P < 0.05; **** P < 0.0001, Student t-test, using GraphPad Prism.

Article Snippet: Membranes were incubated with primary antibodies against Notch1 (D1E11), Notch1-IC (Val1744:D3B8) (Cell Signaling), GAPDH, and β-tubulin (Santa Cruz Biotechnology).

Techniques: Injection, Immunohistochemistry, Flow Cytometry, Marker

Journal: Frontiers in Immunology

Article Title: Sulindac sulfide as a non-immune suppressive γ-secretase modulator to target triple-negative breast cancer

doi: 10.3389/fimmu.2023.1244159

Figure Lengend Snippet: SS does not suppress T-cell proliferation, but blocks BM-MDSC-mediated immune-suppressive activity. T-cells (CD3 + ) were isolated from naïve FVB (female) mice using a negative T cells isolation kit (Stemcell Technologies). Isolated T cells were then labeled with 1 μM CFSE and plated on 24-well culture plates coated with α-CD3 and α-CD28 (1 μg/ml each). T cells were treated with SS (5, 25 or 50 μM SS) at the beginning of incubation, and T-cells proliferation was measured after 72 hours by CFSE dilution using Flow Cytometry (A) . Isolated T-cells from naïve FVB (female) were cultured with plate-bound anti-CD3 and anti-CD28 and were treated with SS (5, 25 or 50 μM SS). Following 72 hours of culture, the expression of Cleaved Notch1 (CN1) and Notch1 was measured by Western Blotting (B) , and IL-2 production was assessed by ELISA (C) . Bone marrow cells were harvested from FVB mice and cultured with GCSF, GM-CSG, and IL-6 (20 ng/ml each) for four days to generate bone marrow-derived MDSC (BM-MDSC) in the presence or absence of SS (5, 25 or 50 μM). CFSE labeled T-cells were co-culture with BM-MDSC at a 4:1 (T-cells: MDSC ratio) with SS (5, 25 or 50 μM) on plate-bound anti-CD3 and anti-CD28 (1 μg/ml each) plate. T-cell proliferation was measured after 72 hours by CFSE dilution using Flow Cytometry (D) . Data are means ± SD; P-values: **** P < 0.0001, one-way ANOVA for multiple comparisons, using GraphPad Prism.

Article Snippet: Membranes were incubated with primary antibodies against Notch1 (D1E11), Notch1-IC (Val1744:D3B8) (Cell Signaling), GAPDH, and β-tubulin (Santa Cruz Biotechnology).

Techniques: Activity Assay, Isolation, Labeling, Incubation, Flow Cytometry, Cell Culture, Expressing, Western Blot, Enzyme-linked Immunosorbent Assay, Derivative Assay, Co-Culture Assay

Journal: American Journal of Translational Research

Article Title: Down-regulation of NOTCH1 and PKM2 can inhibit the growth and metastasis of colorectal cancer cells

doi:

Figure Lengend Snippet: Primer sequences

Article Snippet: Afterwards, the sections were subjected to 16-h incubation (4°C) with primary antibodies against PKM2 and NOTCH1 and then 2-h incubation (indoor temperature) with horseradish peroxidase (HRP)-conjugated secondary antibody (Beijing Zhongshan Jinqiao Biological Technology Co., Ltd., Beijing, CN).

Techniques:

Journal: American Journal of Translational Research

Article Title: Down-regulation of NOTCH1 and PKM2 can inhibit the growth and metastasis of colorectal cancer cells

doi:

Figure Lengend Snippet: Immunohistochemistry. (A and B) show positive expression of PKM2 and NOTCH1 in colorectal cancer tissue, while (C and D) show no expression of the two in normal tissue (×200).

Article Snippet: Afterwards, the sections were subjected to 16-h incubation (4°C) with primary antibodies against PKM2 and NOTCH1 and then 2-h incubation (indoor temperature) with horseradish peroxidase (HRP)-conjugated secondary antibody (Beijing Zhongshan Jinqiao Biological Technology Co., Ltd., Beijing, CN).

Techniques: Immunohistochemistry, Expressing

Journal: American Journal of Translational Research

Article Title: Down-regulation of NOTCH1 and PKM2 can inhibit the growth and metastasis of colorectal cancer cells

doi:

Figure Lengend Snippet: Analysis of correlation between PKM2 and NOTCH1 in colorectal cancer tissue.

Article Snippet: Afterwards, the sections were subjected to 16-h incubation (4°C) with primary antibodies against PKM2 and NOTCH1 and then 2-h incubation (indoor temperature) with horseradish peroxidase (HRP)-conjugated secondary antibody (Beijing Zhongshan Jinqiao Biological Technology Co., Ltd., Beijing, CN).

Techniques:

Journal: American Journal of Translational Research

Article Title: Down-regulation of NOTCH1 and PKM2 can inhibit the growth and metastasis of colorectal cancer cells

doi:

Figure Lengend Snippet: Relationship between PKM2 and NOTCH1 expressions and clinical features of colorectal cancer

Article Snippet: Afterwards, the sections were subjected to 16-h incubation (4°C) with primary antibodies against PKM2 and NOTCH1 and then 2-h incubation (indoor temperature) with horseradish peroxidase (HRP)-conjugated secondary antibody (Beijing Zhongshan Jinqiao Biological Technology Co., Ltd., Beijing, CN).

Techniques: Marker

Journal: American Journal of Translational Research

Article Title: Down-regulation of NOTCH1 and PKM2 can inhibit the growth and metastasis of colorectal cancer cells

doi:

Figure Lengend Snippet: Cox regression analysis

Article Snippet: Afterwards, the sections were subjected to 16-h incubation (4°C) with primary antibodies against PKM2 and NOTCH1 and then 2-h incubation (indoor temperature) with horseradish peroxidase (HRP)-conjugated secondary antibody (Beijing Zhongshan Jinqiao Biological Technology Co., Ltd., Beijing, CN).

Techniques:

Journal: American Journal of Translational Research

Article Title: Down-regulation of NOTCH1 and PKM2 can inhibit the growth and metastasis of colorectal cancer cells

doi:

Figure Lengend Snippet: NOTCH1 and PKM2 expressions in colorectal cancer cells. A, B. qRT-PCR was utilized to determine mRNA levels of PKM2 and NOTCH1 in colorectal cancer cells. C-F. Western blot was utilized to determine the protein levels of PKM2 and NOTCH1 in colorectal cancer cells. ***P<0.001; **P<0.01; ****P<0.0001.

Article Snippet: Afterwards, the sections were subjected to 16-h incubation (4°C) with primary antibodies against PKM2 and NOTCH1 and then 2-h incubation (indoor temperature) with horseradish peroxidase (HRP)-conjugated secondary antibody (Beijing Zhongshan Jinqiao Biological Technology Co., Ltd., Beijing, CN).

Techniques: Quantitative RT-PCR, Western Blot

Journal: American Journal of Translational Research

Article Title: Down-regulation of NOTCH1 and PKM2 can inhibit the growth and metastasis of colorectal cancer cells

doi:

Figure Lengend Snippet: PKM2 and NOTCH1 expressions in colorectal cancer cells treated with compounds 3K and TGN. A-D. qRT-PCR was carried out for measuring mRNA levels of PKM2 and NOTCH1 in the 4 groups of colorectal cancer cells after 24-h incubation with DMSO (as a control), compounds 3K, TGN, or the combination of compound 3K and TGN. E-J. Western blot was conducted to determine the protein levels of PKM2 and NOTCH1 in the 4 groups after 24 h incubation. *P<0.05; **P<0.01; ***P<0.001, D, DMSO; P, Compound 3K; N, tangeretin; P+N, Compound 3K+tangeretin.

Article Snippet: Afterwards, the sections were subjected to 16-h incubation (4°C) with primary antibodies against PKM2 and NOTCH1 and then 2-h incubation (indoor temperature) with horseradish peroxidase (HRP)-conjugated secondary antibody (Beijing Zhongshan Jinqiao Biological Technology Co., Ltd., Beijing, CN).

Techniques: Quantitative RT-PCR, Incubation, Control, Western Blot