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: RL95-2 (ATCC CRL-1671, RRID:CVCL_0505), AN3CA (ATCC HTB-111, RRID:CVCL_0028), and KLE (ATCC CRL-1622, RRID:CVCL_1329) EC cell lines were cultured in RPMI 1640 medium (VWR,CAT# 16777-145) supplemented with 10% fetal bovine serum (VWR, cat # 89510-190), 1% penicillin-streptomycin (VWR, cat# 16777-164), and 2 mmol/L L-glutamine (Gibco, cat# 25030081) at 37°C in a humidified atmosphere containing 5% CO2.

Techniques: Activation Assay, Chromatin Immunoprecipitation, Comparison

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: RL95-2 (ATCC CRL-1671, RRID:CVCL_0505), AN3CA (ATCC HTB-111, RRID:CVCL_0028), and KLE (ATCC CRL-1622, RRID:CVCL_1329) EC cell lines were cultured in RPMI 1640 medium (VWR,CAT# 16777-145) supplemented with 10% fetal bovine serum (VWR, cat # 89510-190), 1% penicillin-streptomycin (VWR, cat# 16777-164), and 2 mmol/L L-glutamine (Gibco, cat# 25030081) at 37°C in a humidified atmosphere containing 5% CO2.

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

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 of the AN3CA subcutaneous xenograft study design. Tumor growth was monitored by caliper measurements. (B) Tumor growth curves showing mean ± SEM tumor volume over time. (C) Endpoint tumor volumes at Day 28. Each dot represents an individual mouse. (D) Kaplan–Meier analysis of time to humane tumor endpoint. Statistical significance was determined using mixed-effects model analysis (B), Kruskal–Wallis test with Dunn’s multiple-comparison correction (C), and log-rank test (D). *, P < 0.05; **, P < 0.01; ***, P < 0.001.

Article Snippet: RL95-2 (ATCC CRL-1671, RRID:CVCL_0505), AN3CA (ATCC HTB-111, RRID:CVCL_0028), and KLE (ATCC CRL-1622, RRID:CVCL_1329) EC cell lines were cultured in RPMI 1640 medium (VWR,CAT# 16777-145) supplemented with 10% fetal bovine serum (VWR, cat # 89510-190), 1% penicillin-streptomycin (VWR, cat# 16777-164), and 2 mmol/L L-glutamine (Gibco, cat# 25030081) at 37°C in a humidified atmosphere containing 5% CO2.

Techniques: Comparison