Journal: Cancer Research Communications

Article Title: Auranofin Potentiates Cisplatin Response in the Setting of Context-Dependent NOTCH-Associated Signaling States in Endometrial Cancer

doi: 10.1158/2767-9764.CRC-26-0404

Figure Lengend Snippet: Upper-quartile NOTCH3 copy number is associated with poorer OS in endometrial cancer. A, Kaplan–Meier analysis of OS in the TCGA-UCEC cohort stratified by NOTCH3 copy number using upper- vs. lower-quartile grouping. Tumors with elevated NOTCH3 copynumber exhibited significantly reduced OS compared with tumors harboring low copy number. B, Kaplan–Meier analysis of OS stratified by NOTCH3 RNA expression using upper- vs. lower-quartile grouping. NOTCH3 RNA expression demonstrated only a modest, nonsignificant association with outcomes. C, Kaplan–Meier analysis of OS according to NOTCH3 mutation status. No significant association between NOTCH3 mutation status and OS 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). CNV, copy-number variation.

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

Techniques: Analysis, Low Copy Number, RNA Expression, Mutagenesis

Journal: Cancer Research Communications

Article Title: Auranofin Potentiates Cisplatin Response in the Setting of Context-Dependent NOTCH-Associated Signaling States in Endometrial Cancer

doi: 10.1158/2767-9764.CRC-26-0404

Figure Lengend Snippet: NOTCH3 protein expression and sensitivity to AuR and CDDP in endometrial cancer cell lines. A, Representative Western blot analysis of full-length NOTCH3 (NOTCH3-FL) and a cleaved NOTCH3/NTM-reactive species in endometrial cancer cell lines. GAPDH served as a loading control. B, Summary of AuR and CDDP IC 50 values across endometrial cancer cell lines. IC 50 values are presented as mean ± SEM from independent biological replicates. C, Dose–response analysis of AuR sensitivity in endometrial cancer cell lines. D, Dose–response analysis of CDDP sensitivity in endometrial cancer cell lines. Dose–response curves in C and D were fitted using a four-parameter logistic regression model.

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

Techniques: Expressing, Sensitivity, Western Blot, Analysis, Control

Journal: Cancer Research Communications

Article Title: Auranofin Potentiates Cisplatin Response in the Setting of Context-Dependent NOTCH-Associated Signaling States in Endometrial Cancer

doi: 10.1158/2767-9764.CRC-26-0404

Figure Lengend Snippet: NOTCH3 depletion alters AuR and CDDP response in a context-dependent manner across endometrial cancer models. 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 (left), with replicate-level AUC quantification (right). E, Dose–response analysis of CDDP response in AN3CA cells following NOTCH3 depletion. Representative nonlinear regression curves are shown (left), with replicate-level AUC quantification (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. Ctrl, control; KD, knockdown.

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

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

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

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

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