Journal: eBioMedicine

Article Title: Proteomic profiling and molecular reclassification of high-grade serous ovarian cancer identifies prognostic subtypes and immunotherapy biomarkers

doi: 10.1016/j.ebiom.2026.106248

Figure Lengend Snippet: Proteomic alterations of HGSOC in early stage and advanced stage . A. Kaplan–Meier curves for OS between early and advanced samples (n = 113, early = 17, advanced = 96) (log-rank test). B. PCA of proteins in 17 early samples and 96 advanced samples. Blue, early samples, orange, advanced samples. C. Volcano plots displaying the DEPs in early and advanced samples after applying a 1.5-fold change in expression with p < 0.05 (Wilcoxon rank-sum test). Proteins significantly enriched in the early and advanced samples are represented as blue/orange-filled dots. Orange, upregulation in advanced sample; blue, upregulation in early sample; grey, not significant. D. Comparison of enriched pathways in early and advanced samples. E. Heatmap of DEPs between early and advanced samples. F. Diagram representing the significant proteins in carbon metabolism pathway (enriched in early samples) (top panel) and pathways in cancer (enriched in advanced samples) (bottom panel). G. Kaplan–Meier survival curves for OS based on protein expressions of ACO2, ENO1, PDGFRB, and EGLN1 in HGSOC (log-rank test). H. Quantification of ACO2 expression in human OC tissues compared to normal ovary (NO) tissues, as detected by qPCR (Student's t-test). I. Cell proliferation of OVCAR8-ACO2-OE cells was assessed by CCK-8 assay (Student's t-test). J-K. Glucose uptake (K) and lactate production (L) in the culture medium were measured in OVCAR8-NC and OVCAR8-ACO2-OE cells (Student's t-test). L. Heatmap of PRKCA, PPP3R1, AKT2, and PLCG1 between early and advanced samples. M. Volcano plot indicating the correlation between the expression of PPP3R1 and the prognosis of HGSOC. N. Kaplan–Meier survival curves for OS based on protein expressions of PPP3R1 in patients with HGSOC (log-rank test). O. Western blot experiment showing the expression level of VEGF in OVCAR8 cells after knockdown of PPP3R1. P. Representative images and quantification of the tube formation assay of HUVEC cells. HUVEC were cultured with OVCAR8-siNC, OVCAR8-siPPP3R1#1 or OVCAR8-siPPP3R1#2 cells (Student's t-test). Scale bar = 50 μm. Q. IF staining of VEGFR in HUVEC cells after co-culture with OVCAR8-siNC, OVCAR8-siPPP3R1#1 or OVCAR8-siPPP3R1#2 cells. Scale bar = 10 μm. R. Representative images of EdU (red) and DAPI (blue) staining in OVCAR8 cells transfected with specific siRNAs of PPP3R1. The right graph shows the quantification of the percentage of EdU-positive cells (Student's t-test). Scale bar = 100 μm. S. CCK-8 assays were performed to evaluate the cell proliferation of OVCAR8 cells transfected with specific siRNAs of PPP3R1 (Student's t-test). T. Representative images of migration and invasion of OVCAR8 cells following GOSR2 knockdown. The right panel represent the quantitative analysis of cell numbers per field (Student's t-test). Scale bar = 100 μm. U. mIHC staining of a representative HGSOC tissue section for PRKCA, PPP3R1, AKT2, PLCG1, and GOSR2. Scale bar = 50 μm. V. Quantification of PRKCA, PPP3R1, AKT2, PLCG1, and GOSR2 staining intensity across human normal, early-stage, and advanced-stage OC tissues (ANOVA). ∗∗∗ p < 0.001, ∗∗ p < 0.01, ∗ p < 0.05.

Article Snippet: Proteins were separated using SurePAGE gels and transferred to the PVDF membranes, then the membranes were blocked, incubated in primary antibodies: GOSR2 antibody (Proteintech, 66134-1-Ig, RRID: AB_2881533 ), FGF2 antibody (Abcam, ab92337, RRID: AB_2049652 ), SEC24D antibody (Proteintech, 13673-1-AP, RRID: AB_2877968 ), PPP3R1 antibody (Proteintech, 13210-1-AP, RRID: AB_2252760 ), VEGF antibody (Absin, abs123884, RRID: AB_3096488 ), EGLN1 antibody (Proteintech, 19886-1-AP, RRID: AB_10641986 ), PDGFRB antibody (Proteintech, 13449-1-AP, RRID: AB_2162644 ), ACO2 antibody (ABclonal, A4524, RRID: AB_2863288 ), β-ACTIN antibody (Proteintech, 66009-1-Ig, RRID: AB_2687938 ).

Techniques: Expressing, Comparison, CCK-8 Assay, Western Blot, Knockdown, Tube Formation Assay, Cell Culture, Staining, Co-Culture Assay, Transfection, Migration

Journal: Cancer Research Communications

Article Title: m6A Modification of ATOX1 Inhibits Acute Myeloid Leukemia Progression by Promoting Cuproptosis

doi: 10.1158/2767-9764.CRC-25-0436

Figure Lengend Snippet: ATOX1 knockdown inhibits ES-/Cu-induced cuproptosis in AML cells. A, Cell Counting Kit-8 assay for detecting the viability of AML cells transfected with oe-NC or oe-ATOX1 treated with different concentrations of ES/Cu (0, 2, 10, 50, and 200 nmol/L and ES and CuCl 2 combined in a 1:1 ratio). AML cells transfected with sh-NC or sh- ATOX1 were treated with 200 nmol/L ES/Cu for 72 hours. B, Cell Counting Kit-8 assay for detecting the viability of AML cells. C, EDU staining assay for detecting the cell proliferation of AML cells. Scale bar, 25 μm. D, Flow cytometry for detecting the cell death of AML cells. E, Western blot analysis of lipoylated DLAT expression in AML cells. F, Western blot analysis of Fe–S cluster protein (FDX1, LIAS, and ACO2) expression in AML cells. Data are shown as mean ± SD. n = 3. SSC, side scatter.

Article Snippet: ACO2 , 11134-1-AP , 1:5,000 , Proteintech , AB_2289288.

Techniques: Knockdown, Cell Counting, Transfection, Staining, Flow Cytometry, Western Blot, Expressing

Journal: Cancer Research Communications

Article Title: m6A Modification of ATOX1 Inhibits Acute Myeloid Leukemia Progression by Promoting Cuproptosis

doi: 10.1158/2767-9764.CRC-25-0436

Figure Lengend Snippet: ATOX1 overexpression promotes ES-/Cu-induced cuproptosis in AML cells. AML cells transfected with oe-NC or oe- ATOX1 were treated with 200 nmol/L ES/Cu for 72 hours. A, Cell Counting Kit-8 assay for detecting the viability of AML cells. B, EDU staining assay for detecting the cell proliferation of AML cells. Scale bar, 25 μm. C, Flow cytometry for detecting the cell death of AML cells. D, Western blot analysis of lipoylated DLAT expression in AML cells. E, Western blot analysis of Fe–S cluster protein (FDX1, LIAS, and ACO2) expression in AML cells. F, Cell Counting Kit-8 assay for detecting the viability of AML cells transfected with oe-NC or oe-ATOX1 and sh-NC or sh- FDX1 treated with different concentrations of ES/Cu (0, 2, 10, 50, and 200 nmol/L) for 72 hours. Data are shown as mean ± SD. n = 3. SSC, side scatter.

Article Snippet: ACO2 , 11134-1-AP , 1:5,000 , Proteintech , AB_2289288.

Techniques: Over Expression, Transfection, Cell Counting, Staining, Flow Cytometry, Western Blot, Expressing

Journal: Cancer Research Communications

Article Title: m6A Modification of ATOX1 Inhibits Acute Myeloid Leukemia Progression by Promoting Cuproptosis

doi: 10.1158/2767-9764.CRC-25-0436

Figure Lengend Snippet: ALKBH5-mediated m6A modification regulating ATOX1 expression affects cuproptosis in AML cells. AML cells transfected with oe- ATOX1 and/or oe- ALKBH5 were treated with 200 nmol/L ES/Cu for 72 hours. A, Western blot analysis of ATOX1 and ALKBH5 expression in AML cells. B, CCK-8 assay for detecting the viability of AML cells. C, EDU staining assay for detecting the cell proliferation of AML cells. Scale bar, 25 μm. D, Flow cytometry for detecting the cell death of AML cells. E, Western blot analysis of lipoylated DLAT expression in AML cells. F, Western blot analysis of Fe–S cluster protein (FDX1, LIAS, and ACO2) expression in AML cells. Data are shown as mean ± SD. n = 3. SSC, side scatter.

Article Snippet: ACO2 , 11134-1-AP , 1:5,000 , Proteintech , AB_2289288.

Techniques: Modification, Expressing, Transfection, Western Blot, CCK-8 Assay, Staining, Flow Cytometry