rabbit anti human notch1  (Cell Signaling Technology Inc)

Journal: Frontiers in Oncology

Article Title: HIF-2α regulates proliferation, invasion, and metastasis of hepatocellular carcinoma cells via VEGF/Notch1 signaling axis after insufficient radiofrequency ablation

doi: 10.3389/fonc.2022.998295

Figure Lengend Snippet: Expression of HIF-2α, Notch1, and VEGF in HC and their paracancerous tissues of patients after insufficient RFA as revealed by immunohistochemistry. Brown color indicates positive staining. Magnification 400×.

Article Snippet: Our study incorporated the following reagents/materials at different stages: DMEM medium, fetal bovine serum (Gibco, USA), tetrazolium blue (MTT) powder (Dongguan Science and Technology Biology Company), Transwell chambers, Matrigel matrix glue (Corning, USA), mouse anti-VEGF monoclonal antibody (Proteintech), rabbit anti-human HIF-2α, rabbit anti-human Notch1, rabbit anti-human β-actin (monoclonal antibodies; CST), and PT2385 (MCE).

Techniques: Expressing, Immunohistochemistry, Staining

Journal: Frontiers in Oncology

Article Title: HIF-2α regulates proliferation, invasion, and metastasis of hepatocellular carcinoma cells via VEGF/Notch1 signaling axis after insufficient radiofrequency ablation

doi: 10.3389/fonc.2022.998295

Figure Lengend Snippet: HIF-2α, Notch1, and VEGF were involved in the increased invasion and proliferation of HCC induced by insufficient RFA. (A) Representative photos of HCC after insufficient RFA were detected by a phase contrast microscope and crystal violet staining. Magnification 400×. (B) Detection of cell proliferation by MTT. (C) Quantitation analysis of Transwell assay. (D–F) Detection of the mRNA expression of HIF-2α, Notch1, and VEGF. **P < 0.01, ***P < 0.001.

Article Snippet: Our study incorporated the following reagents/materials at different stages: DMEM medium, fetal bovine serum (Gibco, USA), tetrazolium blue (MTT) powder (Dongguan Science and Technology Biology Company), Transwell chambers, Matrigel matrix glue (Corning, USA), mouse anti-VEGF monoclonal antibody (Proteintech), rabbit anti-human HIF-2α, rabbit anti-human Notch1, rabbit anti-human β-actin (monoclonal antibodies; CST), and PT2385 (MCE).

Techniques: Microscopy, Staining, Quantitation Assay, Transwell Assay, Expressing

Journal: Frontiers in Oncology

Article Title: HIF-2α regulates proliferation, invasion, and metastasis of hepatocellular carcinoma cells via VEGF/Notch1 signaling axis after insufficient radiofrequency ablation

doi: 10.3389/fonc.2022.998295

Figure Lengend Snippet: Inhibition of HIF-2α by PT2385 suppressed the VEGF and Notch1 signaling pathway in HCC after insufficient RFA. (A–C) Detection of mRNA expression by RT-qPCR. (D, E) Detection of protein levels by western blotting. *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: Our study incorporated the following reagents/materials at different stages: DMEM medium, fetal bovine serum (Gibco, USA), tetrazolium blue (MTT) powder (Dongguan Science and Technology Biology Company), Transwell chambers, Matrigel matrix glue (Corning, USA), mouse anti-VEGF monoclonal antibody (Proteintech), rabbit anti-human HIF-2α, rabbit anti-human Notch1, rabbit anti-human β-actin (monoclonal antibodies; CST), and PT2385 (MCE).

Techniques: Inhibition, Expressing, Quantitative RT-PCR, Western Blot

Journal: Biomedicines

Article Title: Analysis of Primary Chronic Lymphocytic Leukemia Cells’ Signaling Pathways

doi: 10.3390/biomedicines12030524

Figure Lengend Snippet: qPCR analysis of CLL samples for the mRNA expression of NOTCH1, DELTEX1, HES1, and AIOLOS. cDNA was prepared from RNA isolated from blood samples of CLL patients and compared with normal CD19 + peripheral B cells. Relative expression levels were normalized with GUS gene expression. Data are expressed as 2 −ΔCT . Mean values and SEM of replicates are shown. The values of the control samples are shown as a solid line.

Article Snippet: Briefly, 5 μg of proteins from each sample were separated using SDS-PAGE on a 10% gel, transferred to a nitrocellulose membrane, blocked in Blotto buffer for one hour, and incubated overnight with primary antibodies: Rabbit anti-human AIOLOS, Rabbit anti-human cleaved NOTCH1, Rabbit anti-human NOTCH1, and Rabbit anti-human β-ACTIN (all from Cell Signaling Technology, Inc., Beverly, MA, USA) [ , ].

Techniques: Expressing, Isolation, Gene Expression, Control

Journal: Biomedicines

Article Title: Analysis of Primary Chronic Lymphocytic Leukemia Cells’ Signaling Pathways

doi: 10.3390/biomedicines12030524

Figure Lengend Snippet: Correlation plot of data from gene expression ( top ) and protein analysis ( bottom ) of CLL samples for NOTCH1, HES1, DELTEX, and AIOLOS. Correlation coefficients and p -values are indicated.

Article Snippet: Briefly, 5 μg of proteins from each sample were separated using SDS-PAGE on a 10% gel, transferred to a nitrocellulose membrane, blocked in Blotto buffer for one hour, and incubated overnight with primary antibodies: Rabbit anti-human AIOLOS, Rabbit anti-human cleaved NOTCH1, Rabbit anti-human NOTCH1, and Rabbit anti-human β-ACTIN (all from Cell Signaling Technology, Inc., Beverly, MA, USA) [ , ].

Techniques: Gene Expression

Journal: Biomedicines

Article Title: Analysis of Primary Chronic Lymphocytic Leukemia Cells’ Signaling Pathways

doi: 10.3390/biomedicines12030524

Figure Lengend Snippet: Gene and protein expression profile of primary leukemia samples in peripheral blood and bone marrow. ( A ) Ten B-CLL cells, collected from peripheral blood (PBL) and bone marrow (BM), were analyzed for NOTCH1, DELTEX1, HES1, and AIOLOS gene expression using qRT-PCR. Ct values were normalized to the expression of the GUSB housekeeping gene, and expressed as relative gene expression. ( B ) The protein expression of NOTCH1, HES1, and AIOLOS was analyzed using flow cytometry after the labelling of cells with the corresponding antibodies.

Article Snippet: Briefly, 5 μg of proteins from each sample were separated using SDS-PAGE on a 10% gel, transferred to a nitrocellulose membrane, blocked in Blotto buffer for one hour, and incubated overnight with primary antibodies: Rabbit anti-human AIOLOS, Rabbit anti-human cleaved NOTCH1, Rabbit anti-human NOTCH1, and Rabbit anti-human β-ACTIN (all from Cell Signaling Technology, Inc., Beverly, MA, USA) [ , ].

Techniques: Expressing, Gene Expression, Quantitative RT-PCR, Flow Cytometry

Journal: Biomedicines

Article Title: Analysis of Primary Chronic Lymphocytic Leukemia Cells’ Signaling Pathways

doi: 10.3390/biomedicines12030524

Figure Lengend Snippet: qPCR analysis of CLL samples for the mRNA expression of NOTCH1, DELTEX1, HES1, and AIOLOS. cDNA was prepared from RNA isolated from blood samples of CLL patients and compared with normal CD19 + peripheral B cells. Relative expression levels were normalized with GUS gene expression. Data are expressed as 2 −ΔCT . Mean values and SEM of replicates are shown. The values of the control samples are shown as a solid line.

Article Snippet: Briefly, 5 μg of proteins from each sample were separated using SDS-PAGE on a 10% gel, transferred to a nitrocellulose membrane, blocked in Blotto buffer for one hour, and incubated overnight with primary antibodies: Rabbit anti-human AIOLOS, Rabbit anti-human cleaved NOTCH1, Rabbit anti-human NOTCH1, and Rabbit anti-human β-ACTIN (all from Cell Signaling Technology, Inc., Beverly, MA, USA) [ , ].

Techniques: Expressing, Isolation, Gene Expression, Control

Journal: Biomedicines

Article Title: Analysis of Primary Chronic Lymphocytic Leukemia Cells’ Signaling Pathways

doi: 10.3390/biomedicines12030524

Figure Lengend Snippet: Correlation plot of data from gene expression ( top ) and protein analysis ( bottom ) of CLL samples for NOTCH1, HES1, DELTEX, and AIOLOS. Correlation coefficients and p -values are indicated.

Article Snippet: Briefly, 5 μg of proteins from each sample were separated using SDS-PAGE on a 10% gel, transferred to a nitrocellulose membrane, blocked in Blotto buffer for one hour, and incubated overnight with primary antibodies: Rabbit anti-human AIOLOS, Rabbit anti-human cleaved NOTCH1, Rabbit anti-human NOTCH1, and Rabbit anti-human β-ACTIN (all from Cell Signaling Technology, Inc., Beverly, MA, USA) [ , ].

Techniques: Gene Expression

Journal: Biomedicines

Article Title: Analysis of Primary Chronic Lymphocytic Leukemia Cells’ Signaling Pathways

doi: 10.3390/biomedicines12030524

Figure Lengend Snippet: Gene and protein expression profile of primary leukemia samples in peripheral blood and bone marrow. ( A ) Ten B-CLL cells, collected from peripheral blood (PBL) and bone marrow (BM), were analyzed for NOTCH1, DELTEX1, HES1, and AIOLOS gene expression using qRT-PCR. Ct values were normalized to the expression of the GUSB housekeeping gene, and expressed as relative gene expression. ( B ) The protein expression of NOTCH1, HES1, and AIOLOS was analyzed using flow cytometry after the labelling of cells with the corresponding antibodies.

Article Snippet: Briefly, 5 μg of proteins from each sample were separated using SDS-PAGE on a 10% gel, transferred to a nitrocellulose membrane, blocked in Blotto buffer for one hour, and incubated overnight with primary antibodies: Rabbit anti-human AIOLOS, Rabbit anti-human cleaved NOTCH1, Rabbit anti-human NOTCH1, and Rabbit anti-human β-ACTIN (all from Cell Signaling Technology, Inc., Beverly, MA, USA) [ , ].

Techniques: Expressing, Gene Expression, Quantitative RT-PCR, Flow Cytometry

Journal: bioRxiv

Article Title: Mutational order and epistasis determine the consequences of FBXW7 mutations during colorectal cancer evolution

doi: 10.1101/2023.08.25.554836

Figure Lengend Snippet: Characterisation of W and F organoids. a. Brightfield microscopy of W and F organoids revealed no observable phenotypic differences. Scale bars represent 500μm. b. Western blot validation showing loss of FBXW7 protein in F organoids. c. Immunofluorescence of W and F organoids with DAPI nuclear stain (blue), F-actin (red), FBXW7 (orange). Scale bars represent 100μm. d. FBXW7 targets the phosphorylated substrates and ubiquitinates these substrates for proteasomal degradation. This included phosphorylated cJun, phosphorylated CCNE1, phosphosylated cMyc, and notch intracellular domain (NICD). The non-phosphorylated substrates were affected to varying degrees. While upregulation of cJun, was observed, there was no change in the quantities of CCNE1, cMYC and NOTCH. e. Volcano plot from bulk RNAseq of F vs W organoids revealed minimal differential expression of genes. Only HLA-DQB1 was found to be significantly downregulated, and GSTM1 significantly upregulated in F organoids. All experiments were performed with N = 3 biological replicates.

Article Snippet: Rabbit anti-human FBXW7 antibody (1:2500, BS-8394R, Bioss, USA), rabbit anti-human phosphor-CJUN antibody (1:2500, PA5-40193, Invitrogen, USA), rabbit anti-human CJUN antibody (1:2500, ab40766, Abcam, USA), rabbit anti-human phosphor-CCNE1 antibody (1:2500, ab52195, Abcam, USA), rabbit anti-human CCNE1 antibody (1:2500, ab33911, Abcam, USA), rabbit anti-human phosphor-CMYC (1:2500, ab185655 and ab185656, Abcam, USA), rabbit anti-human CMYC (1:2500, ab32072, Abcam, USA), rabbit anti-human NICD (1:2500, #4147, Cell signalling technologies, USA), rabbit anti-human NOTCH1 (1:2500, ab52627, Abcam, USA), and rabbit anti-human GAPDH (1:2500, ab52627, Abcam, USA) was used.

Techniques: Microscopy, Western Blot, Biomarker Discovery, Immunofluorescence, Staining, Quantitative Proteomics

Journal: Frontiers in Oncology

Article Title: HIF-2α regulates proliferation, invasion, and metastasis of hepatocellular carcinoma cells via VEGF/Notch1 signaling axis after insufficient radiofrequency ablation

doi: 10.3389/fonc.2022.998295

Figure Lengend Snippet: Expression of HIF-2α, Notch1, and VEGF in HC and their paracancerous tissues of patients after insufficient RFA as revealed by immunohistochemistry. Brown color indicates positive staining. Magnification 400×.

Article Snippet: Our study incorporated the following reagents/materials at different stages: DMEM medium, fetal bovine serum (Gibco, USA), tetrazolium blue (MTT) powder (Dongguan Science and Technology Biology Company), Transwell chambers, Matrigel matrix glue (Corning, USA), mouse anti-VEGF monoclonal antibody (Proteintech), rabbit anti-human HIF-2α, rabbit anti-human Notch1, rabbit anti-human β-actin (monoclonal antibodies; CST), and PT2385 (MCE).

Techniques: Expressing, Immunohistochemistry, Staining

Journal: Frontiers in Oncology

Article Title: HIF-2α regulates proliferation, invasion, and metastasis of hepatocellular carcinoma cells via VEGF/Notch1 signaling axis after insufficient radiofrequency ablation

doi: 10.3389/fonc.2022.998295

Figure Lengend Snippet: HIF-2α, Notch1, and VEGF were involved in the increased invasion and proliferation of HCC induced by insufficient RFA. (A) Representative photos of HCC after insufficient RFA were detected by a phase contrast microscope and crystal violet staining. Magnification 400×. (B) Detection of cell proliferation by MTT. (C) Quantitation analysis of Transwell assay. (D–F) Detection of the mRNA expression of HIF-2α, Notch1, and VEGF. **P < 0.01, ***P < 0.001.

Article Snippet: Our study incorporated the following reagents/materials at different stages: DMEM medium, fetal bovine serum (Gibco, USA), tetrazolium blue (MTT) powder (Dongguan Science and Technology Biology Company), Transwell chambers, Matrigel matrix glue (Corning, USA), mouse anti-VEGF monoclonal antibody (Proteintech), rabbit anti-human HIF-2α, rabbit anti-human Notch1, rabbit anti-human β-actin (monoclonal antibodies; CST), and PT2385 (MCE).

Techniques: Microscopy, Staining, Quantitation Assay, Transwell Assay, Expressing

Journal: Frontiers in Oncology

Article Title: HIF-2α regulates proliferation, invasion, and metastasis of hepatocellular carcinoma cells via VEGF/Notch1 signaling axis after insufficient radiofrequency ablation

doi: 10.3389/fonc.2022.998295

Figure Lengend Snippet: Inhibition of HIF-2α by PT2385 suppressed the VEGF and Notch1 signaling pathway in HCC after insufficient RFA. (A–C) Detection of mRNA expression by RT-qPCR. (D, E) Detection of protein levels by western blotting. *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: Our study incorporated the following reagents/materials at different stages: DMEM medium, fetal bovine serum (Gibco, USA), tetrazolium blue (MTT) powder (Dongguan Science and Technology Biology Company), Transwell chambers, Matrigel matrix glue (Corning, USA), mouse anti-VEGF monoclonal antibody (Proteintech), rabbit anti-human HIF-2α, rabbit anti-human Notch1, rabbit anti-human β-actin (monoclonal antibodies; CST), and PT2385 (MCE).

Techniques: Inhibition, Expressing, Quantitative RT-PCR, Western Blot

Journal: Frontiers in Medicine

Article Title: Podocyte-specific deletion of miR-146a increases podocyte injury and diabetic kidney disease

doi: 10.3389/fmed.2022.897188

Figure Lengend Snippet: Immunofluorescence imaging-based analyses of glomerular sections shows erlotinib administration protects WT and Pod-miR-146a –/– mice from STZ injury via reduction in ErbB4 and EGFR. (A) Representative confocal microscopy images of immunofluorescently labeled glomeruli from WT (top three panels) and Pod-miR146a (bottom three panels) mice treated with vehicle alone (Control), with STZ and vehicle (STZ) or with STZ and erlotinib (STZ Erl). Tissue sections were imaged after staining with DAPI (nuclear marker) and antibodies against ErbB4, EGFR, Notch-1 and Synaptopodin (Synpo, podocyte marker) (as indicated). Scale bar, 50 μm. (B) Bar graphs showing quantification of relative glomerular signal intensity of ErbB4, EGFR and Notch-1 in tissue samples from A. Statistics were performed using two-way ANOVA. Data shown are mean ± SEM ( n = 5/group). * p < 0.05; *** p < 0.001; ns, no significant difference.

Article Snippet: For EGFR, Notch-1 and ErbB4 staining, sections were then incubated with primary antibodies—Rabbit anti-mouse EGFR (Millipore, #06847), mouse anti-mouse ErbB4 (Santa Cruz, #sc-8050) and Rabbit anti-mouse Notch-1 (Rockland, #100-401-407) in the blocking buffer at 4 ° C, overnight.

Techniques: Immunofluorescence, Imaging, Confocal Microscopy, Labeling, Staining, Marker

Journal: Frontiers in Medicine

Article Title: Podocyte-specific deletion of miR-146a increases podocyte injury and diabetic kidney disease

doi: 10.3389/fmed.2022.897188

Figure Lengend Snippet: Mechanistic model. A diagram showing a mechanistic working model. Podocyte expressed miR-146a represses expression of ErbB4 and Notch-1 during homeostatic conditions, thereby controlling the ErbB4/EGFR and TGFβ 1 signaling pathways. Various external stressors or deletion of miR-146a result in de-repression of ErbB4 and Notch-1, thereby driving the harmful ErbB4/EGFR signaling and inducing TGFβ 1. An autocrine feed-forward loop via TGFβ 1 induces the downstream TGFR/Smad3 signaling, that result in podocyte damage, glomerular injury and proteinuria.

Article Snippet: For EGFR, Notch-1 and ErbB4 staining, sections were then incubated with primary antibodies—Rabbit anti-mouse EGFR (Millipore, #06847), mouse anti-mouse ErbB4 (Santa Cruz, #sc-8050) and Rabbit anti-mouse Notch-1 (Rockland, #100-401-407) in the blocking buffer at 4 ° C, overnight.

Techniques: Expressing