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Cell Signaling Technology Inc notch3

Interaction network between ECs and other cell types in the kidneys of LN mice. (A) Heatmap of cell expression correlation between EC subtypes and other cell types. The color denotes the correlation coefficients between the ligand-receptor pair and the cell types. (B) Ligand-receptor relationship between MCs and EC subclusters, when MCs were the receptor cells. (C) Bubble plot showing the expression of Dll4 in EC subclusters. (D) Co-immunolocalization of Dll4 and Pi16 on EC-1, and co-immunolocalization of <t>Notch3</t> and Fn1 on MCs. Scale bar = 50 μm, n = 3.

Notch3, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/anti+notch3/pmc13062793-81-4-5
Average 86 stars, based on 1 article reviews

notch3 - by Bioz Stars, 2026-10

86/100 stars

Images

1) Product Images from "Endothelial cells regulate mesangial cells through the Dll4/Notch3 axis to participate in glomerular injury in lupus nephritis"

Article Title: Endothelial cells regulate mesangial cells through the Dll4/Notch3 axis to participate in glomerular injury in lupus nephritis

Journal: Frontiers in Immunology

doi: 10.3389/fimmu.2026.1720756

Figure Legend Snippet: Interaction network between ECs and other cell types in the kidneys of LN mice. (A) Heatmap of cell expression correlation between EC subtypes and other cell types. The color denotes the correlation coefficients between the ligand-receptor pair and the cell types. (B) Ligand-receptor relationship between MCs and EC subclusters, when MCs were the receptor cells. (C) Bubble plot showing the expression of Dll4 in EC subclusters. (D) Co-immunolocalization of Dll4 and Pi16 on EC-1, and co-immunolocalization of Notch3 and Fn1 on MCs. Scale bar = 50 μm, n = 3.

Techniques Used: Expressing

EC regulate MC via the Dll4/Notch3 axis. (A) ELISA assays were performed to detect the expression of Dll4 in ECs after LPS stimulation. (B) CCK8 assay showing the effects of Dll4 on MC viability. (C) qRT-PCR assay showing Notch3 relative genes were upregulated in MC after Dll4 stimulation. (D) CCK8 assay showing the effects of Tarextumab on MC viability after Dll4 interference. (E) Western blotting analysis of the expression of the Notch3-ICD and its downstream-related proteins HEY1 and HEY2 following interference with Dll4 alone or in combination with Tarextumab. (F) Quantification of Western blotting. Data were representative of three independent experiments (n = 3 per group). (G) Representative images of wound healing assays showing the migration of MC cells treated with Dll4 or Dll4 in combination with Tarextumab. Scale bar = 200 μm. (H) Quantitative comparison of MC cell migration rates among Control, Dll4, and Dll4+Tarextumab treatment groups (n = 3). (I) Transwell analysis of the effect of Dll4 and Tarextumab on MC cell migration. Scale bar = 2 mm for overview. Scale bar = 300 μm for magnified view. (J) Quantitative analysis of the number of migrated cells. Data are presented as mean ± SD (n = 3). *p < 0.05, **p < 0.01. (K) EdU assays were performed to detect the proliferation of MC treated with Dll4 and Tarextumab. Scale bar = 200 μm. (L) Quantitative analysis of MC cell proliferation by EdU assay. Data are presented as mean ± SD (n = 4). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

Figure Legend Snippet: EC regulate MC via the Dll4/Notch3 axis. (A) ELISA assays were performed to detect the expression of Dll4 in ECs after LPS stimulation. (B) CCK8 assay showing the effects of Dll4 on MC viability. (C) qRT-PCR assay showing Notch3 relative genes were upregulated in MC after Dll4 stimulation. (D) CCK8 assay showing the effects of Tarextumab on MC viability after Dll4 interference. (E) Western blotting analysis of the expression of the Notch3-ICD and its downstream-related proteins HEY1 and HEY2 following interference with Dll4 alone or in combination with Tarextumab. (F) Quantification of Western blotting. Data were representative of three independent experiments (n = 3 per group). (G) Representative images of wound healing assays showing the migration of MC cells treated with Dll4 or Dll4 in combination with Tarextumab. Scale bar = 200 μm. (H) Quantitative comparison of MC cell migration rates among Control, Dll4, and Dll4+Tarextumab treatment groups (n = 3). (I) Transwell analysis of the effect of Dll4 and Tarextumab on MC cell migration. Scale bar = 2 mm for overview. Scale bar = 300 μm for magnified view. (J) Quantitative analysis of the number of migrated cells. Data are presented as mean ± SD (n = 3). *p < 0.05, **p < 0.01. (K) EdU assays were performed to detect the proliferation of MC treated with Dll4 and Tarextumab. Scale bar = 200 μm. (L) Quantitative analysis of MC cell proliferation by EdU assay. Data are presented as mean ± SD (n = 4). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

Techniques Used: Enzyme-linked Immunosorbent Assay, Expressing, CCK-8 Assay, Quantitative RT-PCR, Western Blot, Migration, Comparison, Control, EdU Assay

Schematic illustrating the Dll4-Notch3 axis between EC and MC in the glomeruli of LN. In the early stages of LN, EC regulate MC through the Dll4/Notch3 axis to initiate pathogenesis and affect MC proliferation and migration. As the disease progresses, the proliferation and migration of MC damage the structure and function of the glomerulus, aggravating renal injury and leading to glomerulosclerosis and renal fibrosis.

Figure Legend Snippet: Schematic illustrating the Dll4-Notch3 axis between EC and MC in the glomeruli of LN. In the early stages of LN, EC regulate MC through the Dll4/Notch3 axis to initiate pathogenesis and affect MC proliferation and migration. As the disease progresses, the proliferation and migration of MC damage the structure and function of the glomerulus, aggravating renal injury and leading to glomerulosclerosis and renal fibrosis.

Techniques Used: Migration

other:

Article Title: Inhibitors of the notch transcriptional activation complex and methods for use of the same
Article Snippet: The following Ab were used: anti-Notch1val1744 (4147S, Cell Signaling Technology), anti-Notch2val1697 (SAB4502022, Sigma-Aldrich), anti-Notch3 (5276S, Cell Signaling), anti-CSL (5313S, Cell signaling), anti-SP1 (5931S, Cell Signaling), and anti-GAPDH (ab9483 Abcam).

Article Title: Induction of T‐Cell Differentiation by KLF4 in T‐Cell Acute Lymphoblastic Leukemia Cells Harboring Activating Mutation in NOTCH3
Article Snippet: The membranes were probed with the following primary antibodies at a dilution of 1:1000: anti‐GAPDH (0411; Santa Cruz Biotechnology, USA), anti‐KLF4 (#4038; Cell Signaling Technology, USA), and anti‐Notch3 (#2889; Cell Signaling Technology).

Article Title: Synergistic Crosstalk of PACAP and Notch Signaling Pathways in Bone Development
Article Snippet: Anti-Notch3 , rabbit, polyclonal , 1:500 , Cell Signaling, Danvers, MA, USA.

Cell-Signaling:

Article Title: A Patient-Derived Scaffold-Based 3D Culture Platform for Head and Neck Cancer: Preserving Tumor Heterogeneity for Personalized Drug Testing
Article Snippet: .. The primary antibodies used were α-SMA (ab124964, Abcam, Cambridge, UK), anti-BMP4 (ab39973, Abcam), anti-Vimentin (sc-6260, Santa Cruz Biotechnology, Dallas, TX, USA), and anti-NOTCH3 (2889S, Cell Signaling Technology, Danvers, MA, USA). .. The secondary antibodies used were Horseradish peroxidase-conjugated (HRP) anti-rabbit IgG and anti-mouse IgG (both Cell Signaling Technology, Danvers, MA, USA, #7074P2 and #7076P2).

Article Title: A Patient-Derived Scaffold-Based 3D Culture Platform for Head and Neck Cancer: Preserving Tumor Heterogeneity for Personalized Drug Testing
Article Snippet: .. The primary antibodies used were α-SMA (ab124964, Abcam, UK), anti-BMP4 (ab39973, Abcam), anti-Vimentin (sc-6260, Santa Cruz, USA), and anti-NOTCH3 (2889S, Cell Signaling Technology, USA). .. The secondary antibodies used were Horseradish peroxidase-conjugated (HRP) anti-rabbit IgG and anti-mouse IgG (both Cell Signaling Technology, #7074P2 and #7076P2).

Article Title: A Patient-Derived Scaffold-Based 3D Culture Platform for Head and Neck Cancer: Preserving Tumor Heterogeneity for Personalized Drug Testing.
Article Snippet: .. The primary antibodies used were α-SMA (ab124964, Abcam, Cambridge, UK), anti-BMP4 (ab39973, Abcam), anti-Vimentin (sc-6260, Santa Cruz Biotechnology, Dallas, TX, USA), and anti-NOTCH3 (2889S, Cell Signaling Technology, Danvers, MA, USA). .. The secondary antibodies used were Horseradish peroxidase-conjugated (HRP) anti-rabbit IgG and anti-mouse IgG (both Cell Signaling Technology, Danvers, MA, USA, #7074P2 and #7076P2).

Western Blot:

Article Title: MUC1-C auto-regulatory complex with EBNA1 is responsible for latent Epstein-Barr virus-associated gastric cancer progression
Article Snippet: .. Total lysates prepared from non-confluent cells were analyzed by immunoblotting with anti-MUC1-C (16564, 1:1000 dilution; Cell Signaling Technology (CST), Danvers, MA, USA and HM-1630-P1ABX, 1:1000 dilution; Thermo Fisher Scientific, Waltham, MA, USA), anti-β-actin (A5441, 1:5000 dilution; Sigma-Aldrich, Burlington, MA, USA), anti-E2F1 (3742, 1:1000 dilution; CST), anti-EBNA1 (sc-81581, 1:200 dilution; Santa Cruz, Dallas, TX, USA and ab316860, 1:1000 dilution; abcam, Cambridge, MA, USA), anti-DNMT1 (5032, 1:1000 dilution; CST), anti-DNMT3A (3598, 1:1000 dilution; CST), anti-DNMT3B (67259, 1:1000 dilution; CST), anti-PLOD1 (29480-1-AP, 1:100 dilution; Proteintech, Rosemont, IL, USA), anti-E-Cadherin (3195, 1:1000 dilution; CST), anti-CTCF (3418, 1:1000 dilution; CST), anti-RAD21 (ab992, 1:1000 dilution; Abcam), anti-PARP1 (9532, 1:1000; CST), anti-p21 (2947, 1:1000 dilution; CST), anti-survivin (2808, 1:1000 dilution; CST), anti-BCL-XL (2764, 1:1000 dilution; CST), anti-SOX2 (3579, 1:1000 dilution; CST), anti-KLF4 (12173, 1:1000 dilution; CST), anti-MYC (5605, 1:1000 dilution; CST), anti-BZLF1 (sc-53904, 1:500 dilution; Santa Cruz), anti-BMRF1 (sc-58121, 1:500 dilution; Santa Cruz), anti-NOTCH1 (3608, 1:1000 dilution; CST), anti-NOTCH2 (5732, 1:1000 dilution; CST), anti-NOTCH3 (5276, 1:1000 dilution; CST), anti-HEY1 (19929-1-AP, 1:2,000 dilution; Proteintech), anti-α-Tubulin (2144, 1:1000 dilution; CST), anti-VDAC (ab15895; 1:1000 dilution; Abcam), Lamin B1 (66095-1-Ig, 1:1000 dilution; Proteintech) and anti-Histone H3 (ab1791, 1:5000 dilution; Abcam). .. Cells were harvested with 0.05% TrypLE (Gibco, Waltham, MA, USA), washed with PBS.

Incubation:

Article Title: Effect of Notch1 signaling on muscle engraftment and maturation from pluripotent stem cells
Article Snippet: Following washing, cells were resuspended in MACS buffer (BSA, EDTA and 0.09% azide (Miltenyi Biotec)), and the RFP+CD31-CD45ITGA7+VCAM-1+ cell fraction sorted using a FACSAria II. .. Next, membranes were incubated overnight at 4°C with primary antibodies: anti-NOTCH1 (1:1000; D6F11; Cell Signaling), anti-NOTCH2 (1:1000; D76A6; Cell Signaling), anti-NOTCH3 (1:1000; D11B8; Cell Signaling) and anti- α/βTUBULIN (1:1000; 2148, Cell Signaling). .. Data were analyzed using FlowJo software (BD Biosciences).

Image Search Results

A Correlation analysis between NOTCH3 and CD31 mRNA expression based on the cBioPortal. (Spearman r = 0.66, P = 2.57e-59; Pearson r = 0.68, P = 7.99e-63). B The overall survival analysis and C relapse-free survival analysis of CRC patients based on the data from the TCGA database using the Kaplan-Meier Plotter. D GSVA and E GSEA enrichment analysis of CMS4 and CMS others in CRC patients from the TCGA dataset. F GSVA enrichment analysis and G GSEA enrichment analysis of NOTCH3 high and NOTCH3 low groups in CRC patients from the TCGA dataset. H Representative multiplexed immunofluorescence images of NOTCH3 high and NOTCH3 low tissues. Scale bar, 100 μm. I Histogram of MVD in NOTCH3 high and NOTCH3 low samples (Student’s t- test, n = 15–16). J Correlation between NOTCH3-positive area and MVD (Pearson r = 0.4259, P = 0.02). Data are presented as mean ± SD. Statistical significance was determined by the log-rank test ( B , C ) and Student’s t- test (I). * P < 0.05.

Journal: Cell Death & Disease

Article Title: NICD3 mediates pro-angiogenic effects through SMAD3/TGFBI axis in colorectal cancer

doi: 10.1038/s41419-026-08939-z

Figure Lengend Snippet: A Correlation analysis between NOTCH3 and CD31 mRNA expression based on the cBioPortal. (Spearman r = 0.66, P = 2.57e-59; Pearson r = 0.68, P = 7.99e-63). B The overall survival analysis and C relapse-free survival analysis of CRC patients based on the data from the TCGA database using the Kaplan-Meier Plotter. D GSVA and E GSEA enrichment analysis of CMS4 and CMS others in CRC patients from the TCGA dataset. F GSVA enrichment analysis and G GSEA enrichment analysis of NOTCH3 high and NOTCH3 low groups in CRC patients from the TCGA dataset. H Representative multiplexed immunofluorescence images of NOTCH3 high and NOTCH3 low tissues. Scale bar, 100 μm. I Histogram of MVD in NOTCH3 high and NOTCH3 low samples (Student’s t- test, n = 15–16). J Correlation between NOTCH3-positive area and MVD (Pearson r = 0.4259, P = 0.02). Data are presented as mean ± SD. Statistical significance was determined by the log-rank test ( B , C ) and Student’s t- test (I). * P < 0.05.

Article Snippet: Primary antibodies against NOTCH3 (Proteintech, 55114-1-AP) and CD31(Abclonal, A19014) were applied and incubated overnight at 4 °C.

Techniques: Expressing, Immunofluorescence

A Volcano plot showing differentially expressed genes between control and NICD3-overexpressing HCT116 cells using RNA sequencing ( n = 3). B The relative expression of TGFBI mRNA was detected in control and NICD3-overexpressing CRC cells ( n = 3). C The relative expression of TGFBI mRNA was detected in control and NOTCH3-silencing CRC cells ( n = 4). D Western blot analysis was performed to measure the levels of TGFBI in control and NICD3-overexpressing CRC cells. E TGFBI protein concentration in CM from control and NICD3-overexpressing cells was measured by ELISA ( n = 3–5). F TGFBI protein concentration in CM from control, NICD3-overexpressing, and NICD3-overexpressing followed by NOTCH3 silencing HCT116 cells was measured by ELISA ( n = 3). G Representative immunofluorescence images (left panel) and quantification of TGFBI expression in xenografts from control and NICD3-overexpressing CRC cells ( n = 3, right panel). Scale bar, 50 μm. H Representative images of HUVEC tube formation and migration with CM from the control and rTGFBI-treated (50 μg/mL) HUVEC (left panel). Quantitative analysis of the total branching length and migrated cells ( n = 5–6, right panel). Scale bar, 50 μm. I Representative images of HUVEC tube formation with CM from the control and TGFBI silencing CRC cells (left panel). Quantitative analysis of the total branching length ( n = 6, right panel). Scale bar, 50 μm. J Representative images of HUVEC migration with CM from the control and TGFBI silencing CRC cells (left panel). Quantitative analysis of migrated cells ( n = 6, right panel). Scale bar, 50 μm. Data are presented as mean ± SD. Statistical significance was determined by Student’s t- test ( B , C , E , G , H ) and one-way ANOVA ( F , I , J ). * P < 0.05, ** P < 0.01, *** P < 0.001.

Journal: Cell Death & Disease

Article Title: NICD3 mediates pro-angiogenic effects through SMAD3/TGFBI axis in colorectal cancer

doi: 10.1038/s41419-026-08939-z

Figure Lengend Snippet: A Volcano plot showing differentially expressed genes between control and NICD3-overexpressing HCT116 cells using RNA sequencing ( n = 3). B The relative expression of TGFBI mRNA was detected in control and NICD3-overexpressing CRC cells ( n = 3). C The relative expression of TGFBI mRNA was detected in control and NOTCH3-silencing CRC cells ( n = 4). D Western blot analysis was performed to measure the levels of TGFBI in control and NICD3-overexpressing CRC cells. E TGFBI protein concentration in CM from control and NICD3-overexpressing cells was measured by ELISA ( n = 3–5). F TGFBI protein concentration in CM from control, NICD3-overexpressing, and NICD3-overexpressing followed by NOTCH3 silencing HCT116 cells was measured by ELISA ( n = 3). G Representative immunofluorescence images (left panel) and quantification of TGFBI expression in xenografts from control and NICD3-overexpressing CRC cells ( n = 3, right panel). Scale bar, 50 μm. H Representative images of HUVEC tube formation and migration with CM from the control and rTGFBI-treated (50 μg/mL) HUVEC (left panel). Quantitative analysis of the total branching length and migrated cells ( n = 5–6, right panel). Scale bar, 50 μm. I Representative images of HUVEC tube formation with CM from the control and TGFBI silencing CRC cells (left panel). Quantitative analysis of the total branching length ( n = 6, right panel). Scale bar, 50 μm. J Representative images of HUVEC migration with CM from the control and TGFBI silencing CRC cells (left panel). Quantitative analysis of migrated cells ( n = 6, right panel). Scale bar, 50 μm. Data are presented as mean ± SD. Statistical significance was determined by Student’s t- test ( B , C , E , G , H ) and one-way ANOVA ( F , I , J ). * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Primary antibodies against NOTCH3 (Proteintech, 55114-1-AP) and CD31(Abclonal, A19014) were applied and incubated overnight at 4 °C.

Techniques: Control, RNA Sequencing, Expressing, Western Blot, Protein Concentration, Enzyme-linked Immunosorbent Assay, Immunofluorescence, Migration

A Gene Ontology enrichment analysis of NOTCH3 high and NOTCH3 low groups in CRC patients based on the TCGA dataset. B Co-IP assay detected the interaction between NICD3 and SMAD3 in NICD3-overexpressing HCT116 and SW620 cells. C Schematic representation of SMAD3 structural domains. D Structural prediction of the NICD3-SMAD3 complex by AlphaFold3. E Exogenous Co-IP assay detected the interaction between Flag-NICD3 and Myc-SMAD3-MH2 in HEK293T cells. F Schematic illustration of the TGFBI promoter region, indicating the locations of three sets of primers (P1, P2, and P3) relative to the transcription start site. G CUT&RUN-qPCR analysis of SMAD3 recruitment at the TGFBI promoter (P1-P3) in control and NICD3-overexpressing HCT116 cells. Enrichment was calculated as a fold change relative to the Normal Rabbit IgG control ( n = 3). H Luciferase activity of TGFBI promoter in control and NICD3-overexpressing HCT116 cells ( n = 3). Data are presented as mean ± SD. Statistical significance was determined by Student’s t- test ( G , H ). ** P < 0.01; ns not significant.

Journal: Cell Death & Disease

Article Title: NICD3 mediates pro-angiogenic effects through SMAD3/TGFBI axis in colorectal cancer

doi: 10.1038/s41419-026-08939-z

Figure Lengend Snippet: A Gene Ontology enrichment analysis of NOTCH3 high and NOTCH3 low groups in CRC patients based on the TCGA dataset. B Co-IP assay detected the interaction between NICD3 and SMAD3 in NICD3-overexpressing HCT116 and SW620 cells. C Schematic representation of SMAD3 structural domains. D Structural prediction of the NICD3-SMAD3 complex by AlphaFold3. E Exogenous Co-IP assay detected the interaction between Flag-NICD3 and Myc-SMAD3-MH2 in HEK293T cells. F Schematic illustration of the TGFBI promoter region, indicating the locations of three sets of primers (P1, P2, and P3) relative to the transcription start site. G CUT&RUN-qPCR analysis of SMAD3 recruitment at the TGFBI promoter (P1-P3) in control and NICD3-overexpressing HCT116 cells. Enrichment was calculated as a fold change relative to the Normal Rabbit IgG control ( n = 3). H Luciferase activity of TGFBI promoter in control and NICD3-overexpressing HCT116 cells ( n = 3). Data are presented as mean ± SD. Statistical significance was determined by Student’s t- test ( G , H ). ** P < 0.01; ns not significant.

Article Snippet: Primary antibodies against NOTCH3 (Proteintech, 55114-1-AP) and CD31(Abclonal, A19014) were applied and incubated overnight at 4 °C.

Techniques: Co-Immunoprecipitation Assay, Structural Proteomics, Control, Luciferase, Activity Assay

(A) Kaplan–Meier analysis of overall survival in the TCGA-UCEC cohort stratified by NOTCH3 copy-number using upper-versus-lower quartile grouping. Tumors with elevated NOTCH3 copy-number exhibited significantly reduced overall survival compared with tumors harboring low copy-number. (B) Kaplan–Meier analysis of overall survival stratified by NOTCH3 RNA expression using upper-versus-lower quartile grouping. NOTCH3 RNA expression demonstrated only a modest, nonsignificant association with outcome. (C) Kaplan–Meier analysis of overall survival according to NOTCH3 mutation status. No significant association between NOTCH3 mutation status and overall survival was observed. Log-rank P values are indicated on each panel. Tables beneath Kaplan–Meier plots indicate the number of patients at risk at the indicated time points (days).

Journal: bioRxiv

Article Title: Auranofin potentiates cisplatin response through context-dependent NOTCH-associated signaling states in endometrial cancer

doi: 10.64898/2026.05.27.728338

Figure Lengend Snippet: (A) Kaplan–Meier analysis of overall survival in the TCGA-UCEC cohort stratified by NOTCH3 copy-number using upper-versus-lower quartile grouping. Tumors with elevated NOTCH3 copy-number exhibited significantly reduced overall survival compared with tumors harboring low copy-number. (B) Kaplan–Meier analysis of overall survival stratified by NOTCH3 RNA expression using upper-versus-lower quartile grouping. NOTCH3 RNA expression demonstrated only a modest, nonsignificant association with outcome. (C) Kaplan–Meier analysis of overall survival according to NOTCH3 mutation status. No significant association between NOTCH3 mutation status and overall survival was observed. Log-rank P values are indicated on each panel. Tables beneath Kaplan–Meier plots indicate the number of patients at risk at the indicated time points (days).

Article Snippet: Rabbit polyclonal anti-NOTCH3 antibody was from ABclonal Technology (Cat# A13522, RRID:AB_2760384; 1:2,000).

Techniques: Low Copy Number, RNA Expression, Mutagenesis

(A) Western blot analysis of full-length NOTCH3 (NOTCH3-FL) and activated NOTCH3 intracellular domain (NICD3) in EC cell lines. GAPDH served as a loading control. (B) Summary of AuR and CDDP IC50 values across EC cell lines. IC50 values are presented as mean ± SEM. NICD3 abundance was categorized qualitatively based on immunoblot analysis shown in panel A. (C) Dose-response analysis of AuR sensitivity in EC cell lines. (D) Dose-response analysis of CDDP sensitivity in EC cell lines. Dose-response curves in panels C and D were fitted using a four-parameter logistic regression model.

Journal: bioRxiv

Article Title: Auranofin potentiates cisplatin response through context-dependent NOTCH-associated signaling states in endometrial cancer

doi: 10.64898/2026.05.27.728338

Figure Lengend Snippet: (A) Western blot analysis of full-length NOTCH3 (NOTCH3-FL) and activated NOTCH3 intracellular domain (NICD3) in EC cell lines. GAPDH served as a loading control. (B) Summary of AuR and CDDP IC50 values across EC cell lines. IC50 values are presented as mean ± SEM. NICD3 abundance was categorized qualitatively based on immunoblot analysis shown in panel A. (C) Dose-response analysis of AuR sensitivity in EC cell lines. (D) Dose-response analysis of CDDP sensitivity in EC cell lines. Dose-response curves in panels C and D were fitted using a four-parameter logistic regression model.

Article Snippet: Rabbit polyclonal anti-NOTCH3 antibody was from ABclonal Technology (Cat# A13522, RRID:AB_2760384; 1:2,000).

Techniques: Western Blot, Control

(A) Schematic model illustrating canonical NOTCH3 signaling and the proposed mechanism of AuR action. Following ligand-dependent NOTCH3 activation, NICD3 forms a transcriptional activation complex with RBPJ at canonical NOTCH-responsive promoters, including HES1 and HES4 . AuR reduces RBPJ occupancy at canonical NOTCH-responsive promoters. (B–D) Representative RBPJ chromatin immunoprecipitation (ChIP) assays in RL95-2 (B), ARK-1 (C), and AN3CA (D) cells following treatment with DMSO or 2.5 µM AuR. RBPJ occupancy at the HES1 and HES4 loci was measured by qPCR and normalized to input chromatin. Bead-only controls were included to assess nonspecific background enrichment. (E) Representative nonlinear regression curves from one experiment showing CDDP response in AN3CA cells treated with increasing concentrations of CDDP in the absence or presence of 0.75 or 1.0 µM AuR. Relative proliferation was normalized to untreated controls. (F) Replicate-level quantification of relative CDDP IC50 values from independent biological replicates across AuR concentrations, including 0.75, 1.0, and 1.5 µM AuR conditions. IC50 values were normalized to the no-AuR condition. Data are presented as mean ± SEM. Statistical significance in panel F was determined using one-way ANOVA with multiple-comparison correction. ns, not significant; *, P < 0.05; **, P < 0.01; ***, P < 0.001.

Journal: bioRxiv

Article Title: Auranofin potentiates cisplatin response through context-dependent NOTCH-associated signaling states in endometrial cancer

doi: 10.64898/2026.05.27.728338

Figure Lengend Snippet: (A) Schematic model illustrating canonical NOTCH3 signaling and the proposed mechanism of AuR action. Following ligand-dependent NOTCH3 activation, NICD3 forms a transcriptional activation complex with RBPJ at canonical NOTCH-responsive promoters, including HES1 and HES4 . AuR reduces RBPJ occupancy at canonical NOTCH-responsive promoters. (B–D) Representative RBPJ chromatin immunoprecipitation (ChIP) assays in RL95-2 (B), ARK-1 (C), and AN3CA (D) cells following treatment with DMSO or 2.5 µM AuR. RBPJ occupancy at the HES1 and HES4 loci was measured by qPCR and normalized to input chromatin. Bead-only controls were included to assess nonspecific background enrichment. (E) Representative nonlinear regression curves from one experiment showing CDDP response in AN3CA cells treated with increasing concentrations of CDDP in the absence or presence of 0.75 or 1.0 µM AuR. Relative proliferation was normalized to untreated controls. (F) Replicate-level quantification of relative CDDP IC50 values from independent biological replicates across AuR concentrations, including 0.75, 1.0, and 1.5 µM AuR conditions. IC50 values were normalized to the no-AuR condition. Data are presented as mean ± SEM. Statistical significance in panel F was determined using one-way ANOVA with multiple-comparison correction. ns, not significant; *, P < 0.05; **, P < 0.01; ***, P < 0.001.

Article Snippet: Rabbit polyclonal anti-NOTCH3 antibody was from ABclonal Technology (Cat# A13522, RRID:AB_2760384; 1:2,000).

Techniques: Activation Assay, Chromatin Immunoprecipitation, Comparison

(A) Western blot validation of NOTCH3 knockdown in AN3CA, KLE, and ARK-1 cells using independent shRNAs targeting NOTCH3 (3A and 3C). GAPDH served as a loading control. (B–D) Dose-response analysis of AuR response in Ctrl KD, NOTCH3A KD, and NOTCH3C KD cells. Representative nonlinear regression curves are shown at left, with replicate-level area under the curve (AUC) quantification shown at right. (E) Dose-response analysis of CDDP response in AN3CA cells following NOTCH3 depletion. Representative nonlinear regression curves are shown at left, with replicate-level AUC quantification shown at right. Relative proliferation values were normalized to vehicle-treated controls. Data are presented as mean ± SEM. AUC values were calculated from replicate dose-response curves. Statistical significance was determined using one-way ANOVA with multiple-comparison correction. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ns, not significant.

Journal: bioRxiv

Article Title: Auranofin potentiates cisplatin response through context-dependent NOTCH-associated signaling states in endometrial cancer

doi: 10.64898/2026.05.27.728338

Figure Lengend Snippet: (A) Western blot validation of NOTCH3 knockdown in AN3CA, KLE, and ARK-1 cells using independent shRNAs targeting NOTCH3 (3A and 3C). GAPDH served as a loading control. (B–D) Dose-response analysis of AuR response in Ctrl KD, NOTCH3A KD, and NOTCH3C KD cells. Representative nonlinear regression curves are shown at left, with replicate-level area under the curve (AUC) quantification shown at right. (E) Dose-response analysis of CDDP response in AN3CA cells following NOTCH3 depletion. Representative nonlinear regression curves are shown at left, with replicate-level AUC quantification shown at right. Relative proliferation values were normalized to vehicle-treated controls. Data are presented as mean ± SEM. AUC values were calculated from replicate dose-response curves. Statistical significance was determined using one-way ANOVA with multiple-comparison correction. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ns, not significant.

Article Snippet: Rabbit polyclonal anti-NOTCH3 antibody was from ABclonal Technology (Cat# A13522, RRID:AB_2760384; 1:2,000).

Techniques: Western Blot, Biomarker Discovery, Knockdown, Control, Comparison

Journal: Frontiers in Immunology

Article Title: Endothelial cells regulate mesangial cells through the Dll4/Notch3 axis to participate in glomerular injury in lupus nephritis

doi: 10.3389/fimmu.2026.1720756

Figure Lengend Snippet: Interaction network between ECs and other cell types in the kidneys of LN mice. (A) Heatmap of cell expression correlation between EC subtypes and other cell types. The color denotes the correlation coefficients between the ligand-receptor pair and the cell types. (B) Ligand-receptor relationship between MCs and EC subclusters, when MCs were the receptor cells. (C) Bubble plot showing the expression of Dll4 in EC subclusters. (D) Co-immunolocalization of Dll4 and Pi16 on EC-1, and co-immunolocalization of Notch3 and Fn1 on MCs. Scale bar = 50 μm, n = 3.

Article Snippet: Primary antibodies used included: Notch3 (CST, 5276S, 1:1000), Hey1 (Proteintech, 19929-1-AP, 1:1000), Hey2 (Proteintech, 105791-1-AP, 1:1000), and β-tubulin (CST, #2128S, 1:1000).

Techniques: Expressing

Journal: Frontiers in Immunology

Article Title: Endothelial cells regulate mesangial cells through the Dll4/Notch3 axis to participate in glomerular injury in lupus nephritis

doi: 10.3389/fimmu.2026.1720756

Figure Lengend Snippet: EC regulate MC via the Dll4/Notch3 axis. (A) ELISA assays were performed to detect the expression of Dll4 in ECs after LPS stimulation. (B) CCK8 assay showing the effects of Dll4 on MC viability. (C) qRT-PCR assay showing Notch3 relative genes were upregulated in MC after Dll4 stimulation. (D) CCK8 assay showing the effects of Tarextumab on MC viability after Dll4 interference. (E) Western blotting analysis of the expression of the Notch3-ICD and its downstream-related proteins HEY1 and HEY2 following interference with Dll4 alone or in combination with Tarextumab. (F) Quantification of Western blotting. Data were representative of three independent experiments (n = 3 per group). (G) Representative images of wound healing assays showing the migration of MC cells treated with Dll4 or Dll4 in combination with Tarextumab. Scale bar = 200 μm. (H) Quantitative comparison of MC cell migration rates among Control, Dll4, and Dll4+Tarextumab treatment groups (n = 3). (I) Transwell analysis of the effect of Dll4 and Tarextumab on MC cell migration. Scale bar = 2 mm for overview. Scale bar = 300 μm for magnified view. (J) Quantitative analysis of the number of migrated cells. Data are presented as mean ± SD (n = 3). *p < 0.05, **p < 0.01. (K) EdU assays were performed to detect the proliferation of MC treated with Dll4 and Tarextumab. Scale bar = 200 μm. (L) Quantitative analysis of MC cell proliferation by EdU assay. Data are presented as mean ± SD (n = 4). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

Techniques: Enzyme-linked Immunosorbent Assay, Expressing, CCK-8 Assay, Quantitative RT-PCR, Western Blot, Migration, Comparison, Control, EdU Assay

Journal: Frontiers in Immunology

Article Title: Endothelial cells regulate mesangial cells through the Dll4/Notch3 axis to participate in glomerular injury in lupus nephritis

doi: 10.3389/fimmu.2026.1720756

Figure Lengend Snippet: Schematic illustrating the Dll4-Notch3 axis between EC and MC in the glomeruli of LN. In the early stages of LN, EC regulate MC through the Dll4/Notch3 axis to initiate pathogenesis and affect MC proliferation and migration. As the disease progresses, the proliferation and migration of MC damage the structure and function of the glomerulus, aggravating renal injury and leading to glomerulosclerosis and renal fibrosis.

Techniques: Migration

Review

notch3 (Cell Signaling Technology Inc)

Structured Review

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