Journal: World Journal of Gastrointestinal Oncology

Article Title: Analysis of the potential biological value of pyruvate dehydrogenase E1 subunit β in human cancer

doi: 10.4251/wjgo.v16.i1.144

Figure Lengend Snippet: The predominant methodical workflow in the current study. PDHB: Pyruvate dehydrogenase E1 subunit β; ROC: Receiver operating characteristic; TMB: Tumor mutation burden; MSI: Microsatellite instability; qRT-RCR: Real-time quantitative PCR.

Article Snippet: The membranes were incubated with primary antibodies of PDHB (Proteintech Group, Wuhan) and β-Tubulin (Boster, Wuhan) overnight on a 4 °C shaker, followed by incubation with secondary antibodies for 1 h, three rinses with TBST rinse solution for 10 min each time and then with an enhanced chemiluminescence kit (Boster, Wuhan) to visualize the blots.

Techniques: Mutagenesis, Real-time Polymerase Chain Reaction

Journal: World Journal of Gastrointestinal Oncology

Article Title: Analysis of the potential biological value of pyruvate dehydrogenase E1 subunit β in human cancer

doi: 10.4251/wjgo.v16.i1.144

Figure Lengend Snippet: Expression levels of pyruvate dehydrogenase E1 subunit β in pan-cancer. A: Comparison of pyruvate dehydrogenase E1 subunit β (PDHB) expression levels in different cancers based on The Cancer Genome Atlas (TCGA) database; B: Expression of PDHB in cancer vs normal tissues in the TCGA database; C: Differential expression of PDHB in cancer and normal tissues in the TCGA joint Genotype Tissue Expression Dataset database; D: Protein levels of PDHB in tumors and normal tissues from the UALCAN database. a P < 0.05, b P < 0.01, c P < 0.001. PDHB: Pyruvate dehydrogenase E1 subunit β; ACC: Adrenocortical carcinoma; BLCA: Bladder urothelial carcinoma; BRCA: Breast invasive carcinoma; CESC: Cervical squamous cell carcinoma and Endocervical adenocarcinoma; CHOL: Cholangiocarcinoma; COAD: Colon adenocarcinoma; DLBC: Lymphoid neoplasm diffuse large B-cell lymphoma; ESCA: Esophageal carcinoma; GBM: Glioblastoma multiforme; HNSC: Head and Neck squamous cell carcinoma; KICH: Kidney chromophobe; KIRC: Kidney renal clear cell carcinoma; KIRP: Kidney renal papillary cell carcinoma; LAML: Acute myeloid leukemia; LGG: Brain lower grade glioma; LIHC: Liver hepatocellular carcinoma; LUAD: Lung adenocarcinoma; LUSC: Lung squamous cell carcinoma; MESO: Mesothelioma; OV: Ovarian serous cystadenocarcinoma; PAAD: Pancreatic adenocarcinoma; PCPG: Pheochromocytoma and Paraganglioma; PRAD: Prostate adenocarcinoma; READ: Rectum adenocarcinoma; SKCM: Skin cutaneous melanoma; STAD: Stomach adenocarcinoma; TGCT: Testicular germ cell tumors; THCA: Thyroid carcinoma; THYM: Thymoma; UCEC: Uterine corpus endometrial carcinoma; UCS: Uterine carcinosarcoma; UVM: Uveal melanoma.

Article Snippet: The membranes were incubated with primary antibodies of PDHB (Proteintech Group, Wuhan) and β-Tubulin (Boster, Wuhan) overnight on a 4 °C shaker, followed by incubation with secondary antibodies for 1 h, three rinses with TBST rinse solution for 10 min each time and then with an enhanced chemiluminescence kit (Boster, Wuhan) to visualize the blots.

Techniques: Expressing, Comparison, Quantitative Proteomics

Journal: World Journal of Gastrointestinal Oncology

Article Title: Analysis of the potential biological value of pyruvate dehydrogenase E1 subunit β in human cancer

doi: 10.4251/wjgo.v16.i1.144

Figure Lengend Snippet: Correlation of pyruvate dehydrogenase E1 subunit β expression with tumor stage in multiple cancers and receiver operating characteristic diagnostic analysis. A: Correlation between pyruvate dehydrogenase E1 subunit β (PDHB) gene expression and tumor staging; B: Receiver operating characteristic curve analysis of PDHB in various cancers (AreaUnderROC > 0.7). ACC: Adrenocortical carcinoma; BLCA: Bladder urothelial carcinoma; BRCA: Breast invasive carcinoma; CHOL: Cholangiocarcinoma; COAD: Colon adenocarcinoma; ESCA: Esophageal carcinoma; HNSC: Head and Neck squamous cell carcinoma; KICH: Kidney chromophobe; KIRC: Kidney renal clear cell carcinoma; KIRP: Kidney renal papillary cell carcinoma; LIHC: Liver hepatocellular carcinoma; LUAD: Lung adenocarcinoma; LUSC: Lung squamous cell carcinoma; MESO: Mesothelioma; PAAD: Pancreatic adenocarcinoma; PRAD: Prostate adenocarcinoma; SKCM: Skin cutaneous melanoma; STAD: Stomach adenocarcinoma; TGCT: Testicular germ cell tumors; THCA: Thyroid carcinoma; UVM: Uveal melanoma.

Article Snippet: The membranes were incubated with primary antibodies of PDHB (Proteintech Group, Wuhan) and β-Tubulin (Boster, Wuhan) overnight on a 4 °C shaker, followed by incubation with secondary antibodies for 1 h, three rinses with TBST rinse solution for 10 min each time and then with an enhanced chemiluminescence kit (Boster, Wuhan) to visualize the blots.

Techniques: Expressing, Diagnostic Assay, Gene Expression

Journal: World Journal of Gastrointestinal Oncology

Article Title: Analysis of the potential biological value of pyruvate dehydrogenase E1 subunit β in human cancer

doi: 10.4251/wjgo.v16.i1.144

Figure Lengend Snippet: Mutation frequency of pyruvate dehydrogenase E1 subunit β in pan-cancer. A: Somatic mutation analysis of pyruvate dehydrogenase E1 subunit β (PDHB) in different cancers; B: CBioPortal shows the mutation type and mutation frequency of PDHB sequences.

Article Snippet: The membranes were incubated with primary antibodies of PDHB (Proteintech Group, Wuhan) and β-Tubulin (Boster, Wuhan) overnight on a 4 °C shaker, followed by incubation with secondary antibodies for 1 h, three rinses with TBST rinse solution for 10 min each time and then with an enhanced chemiluminescence kit (Boster, Wuhan) to visualize the blots.

Techniques: Mutagenesis

Journal: World Journal of Gastrointestinal Oncology

Article Title: Analysis of the potential biological value of pyruvate dehydrogenase E1 subunit β in human cancer

doi: 10.4251/wjgo.v16.i1.144

Figure Lengend Snippet: Correlation analysis of pyruvate dehydrogenase E1 subunit β gene expression with DNA methylation levels in tumor mutation burden, microsatellite instability and pan-cancer. A: Radar plot demonstrating the relationship between tumor mutation burden and pyruvate dehydrogenase E1 subunit β (PDHB) gene expression in various malignancies. Correlation coefficients are indicated by red curves and ranges are indicated by blue values; B: Radar plot showing the relationship between microsatellite instability and PDHB gene expression and various malignancies. Correlation coefficients are shown by blue curves and ranges are shown by green values; C: PDHB methylation levels in the UALCAN database on different cancers. a P < 0.05, b P < 0.01, c P < 0.001. ACC: Adrenocortical carcinoma; BLCA: Bladder urothelial carcinoma; BRCA: Breast invasive carcinoma; CESC: Cervical squamous cell carcinoma and Endocervical adenocarcinoma; CHOL: Cholangiocarcinoma; COAD: Colon adenocarcinoma; DLBC: Lymphoid neoplasm diffuse large B-cell lymphoma; ESCA: Esophageal carcinoma; GBM: Glioblastoma multiforme; HNSC: Head and Neck squamous cell carcinoma; KICH: Kidney chromophobe; KIRC: Kidney renal clear cell carcinoma; KIRP: Kidney renal papillary cell carcinoma; LAML: Acute myeloid leukemia; LGG: Brain lower grade glioma; LIHC: Liver hepatocellular carcinoma; LUAD: Lung adenocarcinoma; LUSC: Lung squamous cell carcinoma; MESO: Mesothelioma; OV: Ovarian serous cystadenocarcinoma; PAAD: Pancreatic adenocarcinoma; PCPG: Pheochromocytoma and Paraganglioma; PRAD: Prostate adenocarcinoma; READ: Rectum adenocarcinoma; SKCM: Skin cutaneous melanoma; STAD: Stomach adenocarcinoma; TGCT: Testicular germ cell tumors; THCA: Thyroid carcinoma; THYM: Thymoma; UCEC: Uterine corpus endometrial carcinoma; UCS: Uterine carcinosarcoma; UVM: Uveal melanoma.

Article Snippet: The membranes were incubated with primary antibodies of PDHB (Proteintech Group, Wuhan) and β-Tubulin (Boster, Wuhan) overnight on a 4 °C shaker, followed by incubation with secondary antibodies for 1 h, three rinses with TBST rinse solution for 10 min each time and then with an enhanced chemiluminescence kit (Boster, Wuhan) to visualize the blots.

Techniques: Gene Expression, DNA Methylation Assay, Mutagenesis, Methylation

Journal: World Journal of Gastrointestinal Oncology

Article Title: Analysis of the potential biological value of pyruvate dehydrogenase E1 subunit β in human cancer

doi: 10.4251/wjgo.v16.i1.144

Figure Lengend Snippet: Correlation of pyruvate dehydrogenase E1 subunit β gene expression with stromal and immune scores in different cancers. A: Correlation analysis of pyruvate dehydrogenase E1 subunit β (PDHB) gene expression with immune scores of bladder urothelial carcinoma (BLCA), breast invasive carcinoma (BRCA), brain lower grade glioma (LGG), lung adenocarcinoma (LUAD), lung squamous cell carcinoma, pancreatic adenocarcinoma (PAAD), thyroid carcinoma (THCA), uterine corpus endometrial carcinoma (UCEC); B: Correlation analysis of PDHB gene expression with stromal scores of BLCA, BRCA, LGG, liver hepatocellular carcinoma, LUAD, mesothelioma, ovarian serous cystadenocarcinoma, PAAD, prostate adenocarcinoma, sarcoma, testicular germ cell tumors, THCA, thymoma, UCEC. a P < 0.05, b P < 0.01, c P < 0.001. BRCA: Breast invasive carcinoma; BLCA: Bladder urothelial carcinoma; LGG: Brain lower grade glioma; LUAD: Lung adenocarcinoma; LUSC: Lung squamous cell carcinoma; PAAD: Pancreatic adenocarcinoma; THCA: Thyroid carcinoma; UCEC: Uterine corpus endometrial carcinoma; BRCA: Breast invasive carcinoma; LIHC: Liver hepatocellular carcinoma; MESO: Mesothelioma; OV: Ovarian serous cystadenocarcinoma; PRAD: Prostate adenocarcinoma; SARC: Sarcoma; TGCT: Testicular germ cell tumors; THCA: Thyroid carcinoma; THYM: Thymoma; UCEC: Uterine corpus endometrial carcinoma.

Article Snippet: The membranes were incubated with primary antibodies of PDHB (Proteintech Group, Wuhan) and β-Tubulin (Boster, Wuhan) overnight on a 4 °C shaker, followed by incubation with secondary antibodies for 1 h, three rinses with TBST rinse solution for 10 min each time and then with an enhanced chemiluminescence kit (Boster, Wuhan) to visualize the blots.

Techniques: Gene Expression

Journal: World Journal of Gastrointestinal Oncology

Article Title: Analysis of the potential biological value of pyruvate dehydrogenase E1 subunit β in human cancer

doi: 10.4251/wjgo.v16.i1.144

Figure Lengend Snippet: Correlation analysis of pyruvate dehydrogenase E1 subunit β expression and tumor immunity. A: TIMER method to analyze the correlation between pyruvate dehydrogenase E1 subunit β (PDHB) and immune cell infiltration; B: CIBERSORT method to analyze PDHB correlation with immune cell infiltration; C: Co-expression analysis of PDHB with tumor chemokines; D: Co-expression analysis of PDHB with tumor chemokine receptors; E: Co-expression analysis of PDHB with immune checkpoint gene. a P < 0.05, b P < 0.01, c P < 0.001. PDHB: Pyruvate dehydrogenase E1 subunit β. ACC: Adrenocortical carcinoma; BLCA: Bladder urothelial carcinoma; BRCA: Breast invasive carcinoma; CESC: Cervical squamous cell carcinoma and Endocervical adenocarcinoma; CHOL: Cholangiocarcinoma; COAD: Colon adenocarcinoma; DLBC: Lymphoid neoplasm diffuse large B-cell lymphoma; ESCA: Esophageal carcinoma; GBM: Glioblastoma multiforme; HNSC: Head and Neck squamous cell carcinoma; KICH: Kidney chromophobe; KIRC: Kidney renal clear cell carcinoma; KIRP: Kidney renal papillary cell carcinoma; LAML: Acute myeloid leukemia; LGG: Brain lower grade glioma; LIHC: Liver hepatocellular carcinoma; LUAD: Lung adenocarcinoma; LUSC: Lung squamous cell carcinoma; MESO: Mesothelioma; OV: Ovarian serous cystadenocarcinoma; PAAD: Pancreatic adenocarcinoma; PCPG: Pheochromocytoma and Paraganglioma; PRAD: Prostate adenocarcinoma; READ: Rectum adenocarcinoma; SKCM: Skin cutaneous melanoma; STAD: Stomach adenocarcinoma; TGCT: Testicular germ cell tumors; THCA: Thyroid carcinoma; THYM: Thymoma; UCEC: Uterine corpus endometrial carcinoma; UCS: Uterine carcinosarcoma; UVM: Uveal melanoma.

Article Snippet: The membranes were incubated with primary antibodies of PDHB (Proteintech Group, Wuhan) and β-Tubulin (Boster, Wuhan) overnight on a 4 °C shaker, followed by incubation with secondary antibodies for 1 h, three rinses with TBST rinse solution for 10 min each time and then with an enhanced chemiluminescence kit (Boster, Wuhan) to visualize the blots.

Techniques: Expressing

Journal: World Journal of Gastrointestinal Oncology

Article Title: Analysis of the potential biological value of pyruvate dehydrogenase E1 subunit β in human cancer

doi: 10.4251/wjgo.v16.i1.144

Figure Lengend Snippet: Pyruvate dehydrogenase E1 subunit β expression levels at the single-cell sequencing level. A and B: CancerSEA database demonstrates the correlation of pyruvate dehydrogenase E1 subunit β (PDHB) expression with multiple biological functions in pan-cancer; C: t-Distributed Stochastic Neighbor Embedding plots showing the distribution of PDHB in ovarian serous cystadenocarcinoma, retinoblastoma, and uveal melanoma at the single-cell level. a P < 0.05, b P < 0.01, c P < 0.001. OV: Ovarian serous cystadenocarcinoma; RB: Retinoblastoma; UM: Uveal melanoma.

Article Snippet: The membranes were incubated with primary antibodies of PDHB (Proteintech Group, Wuhan) and β-Tubulin (Boster, Wuhan) overnight on a 4 °C shaker, followed by incubation with secondary antibodies for 1 h, three rinses with TBST rinse solution for 10 min each time and then with an enhanced chemiluminescence kit (Boster, Wuhan) to visualize the blots.

Techniques: Expressing, Sequencing

Journal: World Journal of Gastrointestinal Oncology

Article Title: Analysis of the potential biological value of pyruvate dehydrogenase E1 subunit β in human cancer

doi: 10.4251/wjgo.v16.i1.144

Figure Lengend Snippet: Enrichment analysis of pyruvate dehydrogenase E1 subunit β-associated genes in pan-cancer. A: Interaction network of pyruvate dehydrogenase E1 subunit β (PDHB)-associated biomarkers derived from the BioGRID database; B: GEPIA2.0 showed that PDHB expression was positively correlated with actin related protein 8 (ACTR8), potassium channel tetramerization domain containing 6 (KCTD6), mutl homolog 1 (MLH1), proteasome 26s subunit, non-ATPase 6 (PSMD6), ribonuclease P/MRP subunit p14 (RPP14), and ubiquitin specific peptidase 19 (USP19) genes; C: Heat map showing PDHB expression positively correlated with 6 genes (ACTR8, KCTD6, MLH1, PSMD6, RPP14, USP19); D: Enrichment analysis of PDHB-related genes. PDHB: Pyruvate dehydrogenase E1 subunit β. ACC: Adrenocortical carcinoma; BLCA: Bladder urothelial carcinoma; BRCA: Breast invasive carcinoma; CESC: Cervical squamous cell carcinoma and Endocervical adenocarcinoma; CHOL: Cholangiocarcinoma; COAD: Colon adenocarcinoma; DLBC: Lymphoid neoplasm diffuse large B-cell lymphoma; ESCA: Esophageal carcinoma; GBM: Glioblastoma multiforme; HNSC: Head and Neck squamous cell carcinoma; KICH: Kidney chromophobe; KIRC: Kidney renal clear cell carcinoma; KIRP: Kidney renal papillary cell carcinoma; LAML: Acute myeloid leukemia; LGG: Brain lower grade glioma; LIHC: Liver hepatocellular carcinoma; LUAD: Lung adenocarcinoma; LUSC: Lung squamous cell carcinoma; MESO: Mesothelioma; OV: Ovarian serous cystadenocarcinoma; PAAD: Pancreatic adenocarcinoma; PCPG: Pheochromocytoma and Paraganglioma; PRAD: Prostate adenocarcinoma; READ: Rectum adenocarcinoma; SKCM: Skin cutaneous melanoma; STAD: Stomach adenocarcinoma; TGCT: Testicular germ cell tumors; THCA: Thyroid carcinoma; THYM: Thymoma; UCEC: Uterine corpus endometrial carcinoma; UCS: Uterine carcinosarcoma; UVM: Uveal melanoma.

Article Snippet: The membranes were incubated with primary antibodies of PDHB (Proteintech Group, Wuhan) and β-Tubulin (Boster, Wuhan) overnight on a 4 °C shaker, followed by incubation with secondary antibodies for 1 h, three rinses with TBST rinse solution for 10 min each time and then with an enhanced chemiluminescence kit (Boster, Wuhan) to visualize the blots.

Techniques: Derivative Assay, Expressing, Ubiquitin Proteomics

Journal: World Journal of Gastrointestinal Oncology

Article Title: Analysis of the potential biological value of pyruvate dehydrogenase E1 subunit β in human cancer

doi: 10.4251/wjgo.v16.i1.144

Figure Lengend Snippet: Drug sensitivity analysis. Correlation of IC 50 with pyruvate dehydrogenase E1 subunit β expression for different drugs. A: Chelerythrine; B: Nelarabine; C: Fludarabine; D: Fenretinide; E: Lapachone; F: Vorinostat; G: Dasatinib; H: Dolastatin 10. PDHB: Pyruvate dehydrogenase E1 subunit β.

Article Snippet: The membranes were incubated with primary antibodies of PDHB (Proteintech Group, Wuhan) and β-Tubulin (Boster, Wuhan) overnight on a 4 °C shaker, followed by incubation with secondary antibodies for 1 h, three rinses with TBST rinse solution for 10 min each time and then with an enhanced chemiluminescence kit (Boster, Wuhan) to visualize the blots.

Techniques: Expressing

Journal: World Journal of Gastrointestinal Oncology

Article Title: Analysis of the potential biological value of pyruvate dehydrogenase E1 subunit β in human cancer

doi: 10.4251/wjgo.v16.i1.144

Figure Lengend Snippet: Results of pyruvate dehydrogenase E1 subunit β expression validation. A: Pyruvate dehydrogenase E1 subunit β (PDHB) expression in the human normal hepatocyte cell line (L-O2) and human hepatoma cell lines (SMMC-7721, HepG2, Huh7, H-97); B: PDHB expression in the human normal gastric mucosal cell line (GES-1) and human gastric cancer cell lines (HGC-27, MGC-803, MKN-45); C: PDHB expression in the human normal colonic epithelial cell line (NCM460) and human colon cancer cell lines (SW620, HCT116); D: PDHB expression in the human normal breast cell line (MCF-10A) and breast cancer cell lines (MDA-MB-231, MCF-7); E: PDHB expression in the human normal prostate cell line (RWPE-2) and prostate cancer cell lines (PC-3, 22Rv1 and DU145); F: Validation of PDHB protein expression in the L-O2 and hepatoma cell lines (HepG2, SMMC-7721, Huh7, H-97); G: Quantitative plots; H: Validation of PDHB protein expression in the GES-1 and gastric cancer cell lines (MKN-45, AGS, HGC-27, MGC-803); I: Quantitative plots. a P < 0.05, b P < 0.01, c P < 0.001. PDHB: Pyruvate dehydrogenase E1 subunit β; L-O2: Human normal hepatocyte cell line; GES-1: Gastric mucosal cells; NS: Not significant.

Article Snippet: The membranes were incubated with primary antibodies of PDHB (Proteintech Group, Wuhan) and β-Tubulin (Boster, Wuhan) overnight on a 4 °C shaker, followed by incubation with secondary antibodies for 1 h, three rinses with TBST rinse solution for 10 min each time and then with an enhanced chemiluminescence kit (Boster, Wuhan) to visualize the blots.

Techniques: Expressing, Biomarker Discovery

Journal: World Journal of Gastrointestinal Oncology

Article Title: Analysis of the potential biological value of pyruvate dehydrogenase E1 subunit β in human cancer

doi: 10.4251/wjgo.v16.i1.144

Figure Lengend Snippet: Effects of siRNA-pyruvate dehydrogenase E1 subunit β on proliferation, migration and invasion of Huh7 hepatoma cells. A: Transfection efficiency of siRNA-pyruvate dehydrogenase E1 subunit β (PDHB) in Huh7 cell lines; B: The relative expression of mRNA reflects the efficiency of siRNA-PDHB transfection; C: siRNA-PDHB transfection efficiency by protein expression level; D: The CCK-8 method detected the proliferation capacity of Huh7 cell lines; E: Colony formation assay to determine the proliferation capacity of Huh7 cell lines; F: The histogram shows the number of colonies formed; G: Cell scratch assay to detect the migration capacity of Huh7 cell lines; H: Histogram of quantification of cell scratch assay results; I: Transwell method was used to detect the migration and invasion capacity of Huh7 cell lines; J: The histogram shows the number of migrating cells; K: The histogram shows the number of invading cells. a P < 0.05, b P < 0.01, c P < 0.001. PDHB: Pyruvate dehydrogenase E1 subunit β.

Article Snippet: The membranes were incubated with primary antibodies of PDHB (Proteintech Group, Wuhan) and β-Tubulin (Boster, Wuhan) overnight on a 4 °C shaker, followed by incubation with secondary antibodies for 1 h, three rinses with TBST rinse solution for 10 min each time and then with an enhanced chemiluminescence kit (Boster, Wuhan) to visualize the blots.

Techniques: Migration, Transfection, Expressing, CCK-8 Assay, Colony Assay, Wound Healing Assay

Journal: Nature Communications

Article Title: Cholangiocarcinoma PDHA1 succinylation suppresses macrophage antigen presentation via alpha-ketoglutaric acid accumulation

doi: 10.1038/s41467-025-58429-7

Figure Lengend Snippet: a Upregulated genes revealed by succinylation proteomic analysis of cholangiocarcinoma and adjacent non-tumor tissues ( n = 10). b Mass spectrometry identified the succinylation modification site K83 on PDHA1. c Quantification of PDHA1 K83Succ immunostaining performed to compare paired samples ( n = 74). d Kaplan–Meier analysis of OS curves based on PDHA1 K83Succ expression. PDHA1 K83Succ expression associated with poor prognosis in CCA patients. e Representative immunofluorescence staining of CCA and adjacent normal tissues ( n = 7). PDHA1 K83Succ (Red), DAPI (Blue). And statistical chart of PDHA1 K83Succ mean gray values. Scale bars: 50 μm. f Schematic illustrating transposon- and CRISPR-Cas9-( p53 )-based injection of vectors into mice via the tail vein to generate tumors resembling human CCA. The transposon-based vector overexpressing Kras G12D can express luciferase. Created in BioRender. Zhang, N. (2025) https://BioRender.com/y78d999 . Bioluminescence images and signal of C57BL/6 mice after plasmid injection on day 7 and day 28 ( n = 10 biologically independent samples). g Front and back representative images of livers from ( f ). Scale bars: 1 cm. h Kaplan–Meier survival curve of C57BL/6 mice treated as in ( f ). i Tumor incidence of C57BL/6 mice treated as in ( f ). j Schematic illustrating transposon- and CRISPR-Cas9-( Pten )-based injection of vectors into mice via the tail vein to generate tumors resembling human CCA. The transposon-based vector overexpressing Kras G12D can express luciferase. Created in BioRender. Zhang, N. (2025) https://BioRender.com/j69i397 . Bioluminescence images and signal of C57BL/6 mice after plasmid injection on day 7 and day 28 ( n = 8 biologically independent samples). k Front and back representative images of livers from ( j ). Scale bars: 1 cm. l Kaplan–Meier survival curve of C57BL/6 mice treated as in ( j ). m Tumor incidence of C57BL/6 mice treated as in ( j ). Data are presented as mean values ± SD, and P values were calculated using two-tailed unpaired Student’s t -tests ( c , e , f , j ) or log-rank test ( d , h , l ). HR hazard ratio, WT wild type. Source data are provided as a Source Data File.

Article Snippet: Anti-phospho-PDHA1-S232 mAb (81491-1-RR, 1:1000 for Western blot) and anti-phospho-PDHA1-S300 mAb (29583-1-AP, 1:1000 for Western blot) are from Proteintech.

Techniques: Mass Spectrometry, Modification, Immunostaining, Expressing, Immunofluorescence, Staining, CRISPR, Injection, Plasmid Preparation, Luciferase, Two Tailed Test

Journal: Nature Communications

Article Title: Cholangiocarcinoma PDHA1 succinylation suppresses macrophage antigen presentation via alpha-ketoglutaric acid accumulation

doi: 10.1038/s41467-025-58429-7

Figure Lengend Snippet: a tSNE (t-Distributed Stochastic Neighbor Embedding) plots of 15,830 high-quality single cells from spontaneous cholangiocarcinoma mice. b Heatmaps displaying marker genes for each cell type. c Histogram showing the proportion histogram of the eight major subtypes WT (left) and K83R (right). d tSNE plots of five different macrophage subgroups. e Dot plots showing the expression of marker genes for each cell type. f Quantitative RT-PCR analysis of mRNA levels of Cd74 , H2-Ab1 , H2-Aa , Cxcl9 , Cxcl10 changes from PDHA1 WT or PDHA1 K83R spontaneous cholangiocarcinoma mouse tumors isolated CD45 + CD11b + F4/80 + macrophages ( n = 3 biologically independent samples). g Flow cytometry analysis of CD45 + CD11b + F4/80 + MHCII + cells expression in mice with PDHA1 WT or PDHA1 K83R after hydrodynamic tail vein injection ( n = 5 biologically independent samples). h Representative mIHC staining of spontaneous cholangiocarcinoma mouse tumors. MHCII (Red), CD86 (Green), PDHA1 K83Succ (Yellow) and DAPI (Blue). Scale bars: 20 μm. i Quantification of PDHA1 K83Succ + and CD86 + MHCII + cells in PDHA1 WT and PDHA1 K83R spontaneous cholangiocarcinoma mouse tumors ( n = 5 biologically independent samples). j Representative mIHC staining of human CCA samples using antibodies against HLA-DR (Red), CD86 (Green), PDHA1 K83Succ (Yellow) and DAPI (Blue). Scale bars: 20 μm. k Pearson correlation between the levels of PDHA1 K83Succ + and CD86 + HLA-DR + in human CCA samples ( n = 20). Data are presented as mean values ± SD, and P values were calculated using two-tailed unpaired Student’s t -tests ( f , g , i ) or two-tailed correlation ( k ). Mat matrix, Macro macrophages, WT wild type. RNA-seq data is in the Genome Sequence Archive database and source data are provided as a Source Data File.

Article Snippet: Anti-phospho-PDHA1-S232 mAb (81491-1-RR, 1:1000 for Western blot) and anti-phospho-PDHA1-S300 mAb (29583-1-AP, 1:1000 for Western blot) are from Proteintech.

Techniques: Marker, Expressing, Quantitative RT-PCR, Isolation, Flow Cytometry, Injection, Staining, Two Tailed Test, RNA Sequencing, Sequencing

Journal: Nature Communications

Article Title: Cholangiocarcinoma PDHA1 succinylation suppresses macrophage antigen presentation via alpha-ketoglutaric acid accumulation

doi: 10.1038/s41467-025-58429-7

Figure Lengend Snippet: a Schematic diagram of THP-1 macrophages and cholangiocarcinoma cells co-culture. Created in BioRender. Zhang, N. (2025) https://BioRender.com/c42h873 . After 24 h, macrophages were used for flow analysis, and CM was used for metabolite detection ( n = 5 biological replicates). b Flow cytometry analysis of HLA-DR + MFI values after co-culturing THP-1 and the CM of QBC939 sg PDHA1 with reconstituted expression of PDHA1 WT or PDHA1 K83R after 24 h ( n = 5 biological replicates). c Heatmap representing metabolite changes in tumor WT CM compared with K83 CM was obtained after 24 h culture of tumor cells ( n = 5 biological replicates). d Schematic diagram of the TCA cycle metabolism in cancer. e Metabolite secretion of TCA cycle metabolic substrates α-KG, succinate, fumarate, malate and citrate changes in CM from QBC939 sg PDHA1 with reconstituted expression of PDHA1 WT or PDHA1 K83R. Analysis was performed after 24 h ( n = 5 biological replicates). f Flow cytometry analysis of HLA-DR + MFI values of THP-1 in co-culture with tumor CM. Analysis was performed after 24 h ( n = 5 biological replicates). g PDH enzyme activity of QBC939 sg PDHA1 with reconstituted expression of PDHA1 WT and PDHA1 K83R ( n = 5 biological replicates). Activity is represented as the rate in ΔOD/min. h Western blot analysis showed the expression of PDHA1 pS293, PDHA1 K83Succ changes in QBC939 sg PDHA1 with reconstituted expression of PDHA1 WT, PDHA1 K83R, PDHA1 S293A and PDHA1 K83R S293A. The samples derive from the same experiment but different gels for PDHA1 pS293, PDHA1 K83Succ, PDHA1 and α-Tubulin were processed in parallel. The experiment was repeated three times with similar results. i PDH enzyme activity of QBC939 sg PDHA1 with reconstituted expression of PDHA1 WT, PDHA1 K83R, PDHA1 S293A and PDHA1 K83R S293A ( n = 5 biological replicates). Activity is represented as the rate in ΔOD/min. j CM α-KG concentration of QBC939 sg PDHA1 with reconstituted expression of PDHA1 WT, PDHA1 K83R, PDHA1 S293A and PDHA1 K83R S293A ( n = 5 biological replicates). Data are presented as mean values ± SD, and P values were calculated using two-tailed unpaired Student’s t -tests. Mac macrophages, CM conditioned medium, MFI mean fluorescence intensity. Source data are provided as a Source Data File.

Article Snippet: Anti-phospho-PDHA1-S232 mAb (81491-1-RR, 1:1000 for Western blot) and anti-phospho-PDHA1-S300 mAb (29583-1-AP, 1:1000 for Western blot) are from Proteintech.

Techniques: Co-Culture Assay, Flow Cytometry, Expressing, Activity Assay, Western Blot, Concentration Assay, Two Tailed Test, Fluorescence

Journal: Nature Communications

Article Title: Cholangiocarcinoma PDHA1 succinylation suppresses macrophage antigen presentation via alpha-ketoglutaric acid accumulation

doi: 10.1038/s41467-025-58429-7

Figure Lengend Snippet: a Gene Ontology (GO) analysis of RNA-seq after 24 h co-culture of tumor CM with or without α-KG (8 μM) and THP-1 cells. b Volcano plot to depict the downregulated HLA-DRA , HLA-DPA1 , and HLA-DQA1 expression changes of immune response observed in the RNA-seq data. c Gene Set Enrichment Analysis (GSEA) indicating that positive regulation of MAPK cascade was significantly upregulated after α-KG accumulation. d Western blot analysis showed the expression of the MAPK pathway and HLA-DR, CD74 changes after 24 h co-culture ( n = 3 biological replicates). The samples derive from the same experiment but different gels for p-ERK, ERK, and HLA-DR, another for CD74 and p-P38, another for p-JNK, JNK, and another for P38 and α-Tubulin were processed in parallel. e Diagram of interaction between α-KG and OXGR1 protein using molecular docking. f Western blot analysis showed the expression of p-ERK, HLA-DR, and CD74 changes after 24 h co-culture ( n = 3 biological replicates). The samples derive from the same experiment but different gels for p-ERK, ERK, and HLA-DR, another for CD74, and another for OXGR1 and α-Tubulin were processed in parallel. g Schematic diagram of THP-1 with or without OXGR1 depletion and tumor CM with α-KG (0, 8 μM). Created in BioRender. Zhang, N. (2025) https://BioRender.com/a10o519 . After 24 h, macrophages were used for flow analysis. h Flow cytometry analysis of HLA-DR + MFI values from data ( g ) ( n = 4 biological replicates). i Schematic diagram of subcutaneous tumor treatment. Created in BioRender. Zhang, N. (2025) https://BioRender.com/j80q160 . And representative images of vehicle, montelukast, α-KG or combined on day 26 ( n = 5 biologically independent samples). j Tumor growth of LTP-C9 with overexpression of PDHA1 WT were injected into C57BL/6 mice ( n = 5 biologically independent samples), with vehicle, montelukast, α-KG or combined treatment, beginning on day 14. k Tumor volume from data ( i ) ( n = 5 biologically independent samples). l Tumor weight from data ( i ) ( n = 5 biologically independent samples). Data are presented as mean values ± SD, and P values were calculated using two-tailed unpaired Student’s t -tests ( d , f , h , k , l ) or 2way ANOVA ( j ). Each Western blot was independently repeated three times. Mac macrophages, CM conditioned medium, MFI mean fluorescence intensity. Source data are provided as a Source Data File.

Article Snippet: Anti-phospho-PDHA1-S232 mAb (81491-1-RR, 1:1000 for Western blot) and anti-phospho-PDHA1-S300 mAb (29583-1-AP, 1:1000 for Western blot) are from Proteintech.

Techniques: RNA Sequencing, Co-Culture Assay, Expressing, Western Blot, Flow Cytometry, Over Expression, Injection, Two Tailed Test, Fluorescence

Journal: Nature Communications

Article Title: Cholangiocarcinoma PDHA1 succinylation suppresses macrophage antigen presentation via alpha-ketoglutaric acid accumulation

doi: 10.1038/s41467-025-58429-7

Figure Lengend Snippet: a Immunofluorescence analysis to detect colocalization of PDHA1 and DLST in HCCC9810 and RBE cells. Scale bars: 20 μm. b Western blot analysis of input and anti-FLAG immunoprecipitates (IPs) derived from HEK293T cells transfected with FLAG-PDHA1 and HA-tagged EV, HAT1, KAT2A, CPT1A, DLST. The samples derive from the same experiment but different gels for Succ-K, another for HA, another for Flag, and another for α-Tubulin were processed in parallel. c Western blot analysis of input and anti-FLAG immunoprecipitates (IPs) derived from HEK293T cells transfected with FLAG-PDHA1-EV, FLAG-PDHA1 WT, FLAG-PDHA1 K83R, and HA-DLST. The samples derive from the same experiment but different gels for Succ-K, another for HA, another for Flag, and another for α-Tubulin were processed in parallel. d Western blot analysis of DLST changes from QBC939 cells. e Western blot analysis of PDHA1 K83Succ, PDHA1 pS293, PDHA1 changes from QBC939 cells after DLST depletion. The samples derive from the same experiment but different gels for PDHA1 K83Succ, PDHA1 pS293, and DLST, another for PDHA1, and another for α-Tubulin were processed in parallel. f PDH enzyme activity of QBC939 cells after DLST depletion ( n = 5 biological replicates). Activity is represented as the rate in ΔOD/min. g CM α-KG concentration of QBC939 cells after DLST depletion ( n = 3 biological replicates). h Flow cytometry analysis of HLA-DR + MFI values after co-culturing the CM of QBC939 cells after DLST depletion and THP-1 after 24 h ( n = 5 biological replicates). i Western blot analysis of PDHA1 pS293 and PDHA1 K83Succ changes from QBC939 sg PDHA1 with reconstituted expression of FLAG-PDHA1 WT and FLAG-PDHA1 K83R with or without DLST. The samples derive from the same experiment but different gels for PDHA1 pS293, PDHA1 K83Succ, and DLST, another for PDHA1, and another for α-Tubulin were processed in parallel. j CM α-KG concentration of QBC939 sg PDHA1 with reconstituted expression of FLAG-PDHA1 WT and Flag-PDHA1 K83R with or without DLST ( n = 5 biological replicates). k Western blot analysis of PDHA1 K83Succ, PDHA1 pS293 changes from QBC939 sgPDHA1 with reconstituted expression of FLAG-PDHA1 WT cell line used CPI-613 (0, 50, 100 μM). The samples derive from the same experiment but different gels for PDHA1 K83Succ, PDHA1 pS293, another for PDHA1 and Actin were processed in parallel. l CM α-KG concentration of QBC939 sg PDHA1 with reconstituted expression of FLAG-PDHA1 WT cell line used CPI-613 (0, 50, 100 μM) ( n = 5 biological replicates). m Flow cytometry analysis of HLA-DR + MFI values after co-culturing the CM of QBC939 sg PDHA1 with reconstituted expression of FLAG-PDHA1 WT cell line used CPI-613 (0, 50, 100 μM) ( n = 5 biological replicates). Data are presented as mean values ± SD, and P values were calculated using two-tailed unpaired Student’s t -tests. Each Western blot was independently repeated three times. CM conditioned medium, MFI mean fluorescence intensity. Source data are provided as a Source Data File.

Article Snippet: Anti-phospho-PDHA1-S232 mAb (81491-1-RR, 1:1000 for Western blot) and anti-phospho-PDHA1-S300 mAb (29583-1-AP, 1:1000 for Western blot) are from Proteintech.

Techniques: Immunofluorescence, Western Blot, Derivative Assay, Transfection, Activity Assay, Concentration Assay, Flow Cytometry, Expressing, Two Tailed Test, Fluorescence

Journal: Nature Communications

Article Title: Cholangiocarcinoma PDHA1 succinylation suppresses macrophage antigen presentation via alpha-ketoglutaric acid accumulation

doi: 10.1038/s41467-025-58429-7

Figure Lengend Snippet: a Schematic diagram of subcutaneous tumor treatment. Created in BioRender. Zhang, N. (2025) https://BioRender.com/g40g507 . b Tumor growth of LTP-C9 with overexpression of PDHA1 WT were injected into C57BL/6 mice ( n = 6 biologically independent samples), with vehicle, CPI-613, Gem+Cis or combined treatment, beginning on day 14, CPI-613 and vehicle were administered daily, Gem+Cis were administered on experimental days (ED) 1, 5, 9, and 13. c Representative images of tumor volumes from vehicle, CPI-613, Gem+Cis or combined treatment on day 30 ( n = 6 biologically independent samples). d Tumor volume and weight from data ( b ) ( n = 6 biologically independent samples). e Schematic diagram of spontaneous tumor treatment. Created in BioRender. Zhang, N. (2025) https://BioRender.com/v16u276 . f Bioluminescence images of C57BL/6 mice after plasmid injection on day 7, and mice on day 28 after administration of vehicle, CPI-613, Gem+Cis or combined treatment. g Bioluminescence signal from data ( e ) ( n = 10 biologically independent samples). h Front and back representative images of livers from ( e ). Scale bars: 1 cm. i Kaplan–Meier survival curve of C57BL/6 mice treated as in ( e ) ( n = 10 biologically independent samples). j Tumor incidence from data ( e ). k Flow cytometry analysis of CD45 + CD11b + F4/80 + MHCII + cells expression of vehicle, CPI-613, Gem+Cis or combined treatment ( n = 5 biologically independent samples). l Flow cytometry analysis of CD45 + CD3 + CD4 + T cells expression of vehicle, CPI-613, Gem+Cis or combined treatment ( n = 5 biologically independent samples). m Flow cytometry analysis of CD45 + CD3 + CD8 + T cells expression of vehicle, CPI-613, Gem+Cis or combined treatment ( n = 5 biologically independent samples). Data are presented as mean values ± SD, and P values were calculated using two-tailed unpaired Student’s t -tests ( d , g , k – m ), 2way ANOVA ( b ) or log-rank test ( i ). Gem gemcitabine, Cis cisplatin. Source data are provided as a Source Data File.

Article Snippet: Anti-phospho-PDHA1-S232 mAb (81491-1-RR, 1:1000 for Western blot) and anti-phospho-PDHA1-S300 mAb (29583-1-AP, 1:1000 for Western blot) are from Proteintech.

Techniques: Over Expression, Injection, Plasmid Preparation, Flow Cytometry, Expressing, Two Tailed Test

Journal: NPJ Precision Oncology

Article Title: Cuproptosis-associated PDHA1 promotes sarcoma progression and immunotherapy responsiveness via the E2F1–PD-L1 axis: a multi-omics and clinical validation study

doi: 10.1038/s41698-026-01298-0

Figure Lengend Snippet: Overview of the study revealing PDHA1 as a key regulator of sarcoma progression and immune evasion.

Article Snippet: To modulate cuproptosis, MG63 cells with stable PDHA1 knockdown or control (sh-PDHA1 and sh-NC) were treated with the copper chelator ammonium tetrathiomolybdate (TTM; MedChemExpress, HY-100276).

Techniques:

Journal: NPJ Precision Oncology

Article Title: Cuproptosis-associated PDHA1 promotes sarcoma progression and immunotherapy responsiveness via the E2F1–PD-L1 axis: a multi-omics and clinical validation study

doi: 10.1038/s41698-026-01298-0

Figure Lengend Snippet: A Cumulative distribution function (CDF) plots for consensus clustering with k = 2–6. B Relative change in the area under the CDF curve for different k-values. C Consensus matrix for clustering at k = 2. D Heatmap of CRG expression across subtypes. E Differential expression of CRGs between C1 and C2 in TCGA-SARC. F Protein-protein interaction network of 12 CRGs, highlighting PDHA1 as a hub. G Pearson correlation matrix of CRG expression. H Friends similarity analysis showing PDHA1 as the most central gene.

Article Snippet: To modulate cuproptosis, MG63 cells with stable PDHA1 knockdown or control (sh-PDHA1 and sh-NC) were treated with the copper chelator ammonium tetrathiomolybdate (TTM; MedChemExpress, HY-100276).

Techniques: Expressing, Quantitative Proteomics

Journal: NPJ Precision Oncology

Article Title: Cuproptosis-associated PDHA1 promotes sarcoma progression and immunotherapy responsiveness via the E2F1–PD-L1 axis: a multi-omics and clinical validation study

doi: 10.1038/s41698-026-01298-0

Figure Lengend Snippet: A PDHA1 expression across GEO, TARGET, and ICGC. B ROC curves for PDHA1 diagnostic value. C Distribution of risk score, survival status, and expression in TCGA. D Kaplan-Meier curve of OS. E Time-dependent ROC curves for 1-, 3-, and 5-year OS. F DSS curve of PDHA1 high and low groups. G OS curve in GSE21257 . H OS curve in GSE17674 .

Article Snippet: To modulate cuproptosis, MG63 cells with stable PDHA1 knockdown or control (sh-PDHA1 and sh-NC) were treated with the copper chelator ammonium tetrathiomolybdate (TTM; MedChemExpress, HY-100276).

Techniques: Expressing, Diagnostic Assay

Journal: NPJ Precision Oncology

Article Title: Cuproptosis-associated PDHA1 promotes sarcoma progression and immunotherapy responsiveness via the E2F1–PD-L1 axis: a multi-omics and clinical validation study

doi: 10.1038/s41698-026-01298-0

Figure Lengend Snippet: A Nomogram integrating PDHA1 and clinical variables. B Calibration curves for 1-, 3-, and 5-year OS. C ROC curves for OS prediction. D Time-dependent AUC curves. E DCA showing clinical benefit of the nomogram.

Article Snippet: To modulate cuproptosis, MG63 cells with stable PDHA1 knockdown or control (sh-PDHA1 and sh-NC) were treated with the copper chelator ammonium tetrathiomolybdate (TTM; MedChemExpress, HY-100276).

Techniques:

Journal: NPJ Precision Oncology

Article Title: Cuproptosis-associated PDHA1 promotes sarcoma progression and immunotherapy responsiveness via the E2F1–PD-L1 axis: a multi-omics and clinical validation study

doi: 10.1038/s41698-026-01298-0

Figure Lengend Snippet: A Correlation between PDHA1 expression levels and infiltration of various immune cells across 33 different tumor types, analyzed using CIBERSORT and ssGSEA algorithms (* P < 0.05). B Differences in immune infiltration between high and low PDHA1 expression groups based on the CIBERSORT algorithm. C Correlation analysis between PDHA1 expression and immune infiltration based on the CIBERSORT algorithm. D Differences in immune infiltration between high and low PDHA1 expression groups based on the ssGSEA algorithm. E Correlation analysis between PDHA1 expression and immune infiltration based on the ssGSEA algorithm. F Differences in ESTIMATE scores between high and low PDHA1 expression groups.

Article Snippet: To modulate cuproptosis, MG63 cells with stable PDHA1 knockdown or control (sh-PDHA1 and sh-NC) were treated with the copper chelator ammonium tetrathiomolybdate (TTM; MedChemExpress, HY-100276).

Techniques: Expressing

Journal: NPJ Precision Oncology

Article Title: Cuproptosis-associated PDHA1 promotes sarcoma progression and immunotherapy responsiveness via the E2F1–PD-L1 axis: a multi-omics and clinical validation study

doi: 10.1038/s41698-026-01298-0

Figure Lengend Snippet: A scRNA-seq clustering of SARC immune cells. B Cell lineage annotations. C Heatmap of gene expression aggregated by cell type. D Dot plot of canonical marker genes. E PDHA1 distribution across cell types. F Cell–cell communication network. G Ligand–receptor interaction bubble plot.

Article Snippet: To modulate cuproptosis, MG63 cells with stable PDHA1 knockdown or control (sh-PDHA1 and sh-NC) were treated with the copper chelator ammonium tetrathiomolybdate (TTM; MedChemExpress, HY-100276).

Techniques: Gene Expression, Marker

Journal: NPJ Precision Oncology

Article Title: Cuproptosis-associated PDHA1 promotes sarcoma progression and immunotherapy responsiveness via the E2F1–PD-L1 axis: a multi-omics and clinical validation study

doi: 10.1038/s41698-026-01298-0

Figure Lengend Snippet: A PDHA1 mRNA levels are significantly higher in sarcoma tissues than in matched adjacent normal tissues ( n = 60). B , C Western blot and quantitative analysis showing increased protein expression of PDHA1, PD-L1 and E2F1 in tumor versus normal tissues. D , E Representative IHC staining and semiquantitative analysis in liposarcoma and synovial sarcoma samples demonstrating upregulation of PDHA1, PD-L1, E2F1 and Ki-67 in tumor tissues. Scale bar: 50 μm. F Nomogram incorporating PDHA1 expression, age, gender and metastasis status to predict 1-, 3- and 5-year overall survival (OS) in sarcoma patients. G Kaplan–Meier analysis indicating that high PDHA1 expression is associated with poor OS. H Time-dependent ROC curves showing the predictive performance of the PDHA1-based model for 1-, 3- and 5-year OS. I Dynamic AUC plot evaluating the discrimination ability of the nomogram over time. J Decision curve analysis (DCA) demonstrating the net clinical benefit of the PDHA1-based nomogram. K – M qPCR and Western blot analyses showing that PDHA1 expression is significantly elevated in human sarcoma cell lines (MG63, SW982, SW872) compared with normal control cells (hFOB1.19, HFLS, HPA-V). Data are presented as mean ± SEM. *** P < 0.001.

Article Snippet: To modulate cuproptosis, MG63 cells with stable PDHA1 knockdown or control (sh-PDHA1 and sh-NC) were treated with the copper chelator ammonium tetrathiomolybdate (TTM; MedChemExpress, HY-100276).

Techniques: Western Blot, Expressing, Immunohistochemistry, Control

Journal: NPJ Precision Oncology

Article Title: Cuproptosis-associated PDHA1 promotes sarcoma progression and immunotherapy responsiveness via the E2F1–PD-L1 axis: a multi-omics and clinical validation study

doi: 10.1038/s41698-026-01298-0

Figure Lengend Snippet: A qRT-PCR analysis showing efficient knockdown of PDHA1 by three independent shRNAs and corresponding downregulation of PD-L1 and E2F1 in MG63 and SW982 cells. B Western blot confirms reduced protein levels of PDHA1, E2F1, and PD-L1 in both cell lines. C , D CCK-8 assays demonstrate significantly impaired proliferation after PDHA1 silencing at 24–72 h. E , F EdU immunofluorescence staining and quantification show decreased DNA synthesis in PDHA1-deficient cells. G , H Wound healing assays reveal reduced migration capacity after PDHA1 knockdown. I , J Transwell migration and invasion assays indicate markedly decreased motility and invasiveness in both MG63 and SW982 cells. K , L Colony formation assays demonstrate a substantial reduction in clonogenic potential upon PDHA1 depletion.

Article Snippet: To modulate cuproptosis, MG63 cells with stable PDHA1 knockdown or control (sh-PDHA1 and sh-NC) were treated with the copper chelator ammonium tetrathiomolybdate (TTM; MedChemExpress, HY-100276).

Techniques: Quantitative RT-PCR, Knockdown, Western Blot, CCK-8 Assay, Immunofluorescence, Staining, DNA Synthesis, Migration

Journal: NPJ Precision Oncology

Article Title: Cuproptosis-associated PDHA1 promotes sarcoma progression and immunotherapy responsiveness via the E2F1–PD-L1 axis: a multi-omics and clinical validation study

doi: 10.1038/s41698-026-01298-0

Figure Lengend Snippet: A – C 3D spheroid assays in MG63 cells show significantly reduced spheroid volume and Matrigel invasion area following PDHA1 knockdown over 7 days. D – F In vivo xenograft models demonstrate markedly suppressed tumor growth and decreased tumor weight in the sh-PDHA1 group. G Representative H&E, TUNEL, and IHC staining of xenograft tumors show decreased expression of PDHA1, Ki-67, Cyclin D1, PD-L1, E2F1, Bcl-2, and MMP-9, and increased cleaved Caspase-3 and apoptosis in the sh-PDHA1 group. H Quantification of TUNEL and IHC staining. I , J Western blot and densitometric analysis confirm significant downregulation of PDHA1, PD-L1, E2F1, Bcl-2, and MMP-9, and upregulation of cleaved Caspase-3 in tumor lysates upon PDHA1 knockdown.

Article Snippet: To modulate cuproptosis, MG63 cells with stable PDHA1 knockdown or control (sh-PDHA1 and sh-NC) were treated with the copper chelator ammonium tetrathiomolybdate (TTM; MedChemExpress, HY-100276).

Techniques: Knockdown, In Vivo, TUNEL Assay, Immunohistochemistry, Expressing, Western Blot

Journal: NPJ Precision Oncology

Article Title: Cuproptosis-associated PDHA1 promotes sarcoma progression and immunotherapy responsiveness via the E2F1–PD-L1 axis: a multi-omics and clinical validation study

doi: 10.1038/s41698-026-01298-0

Figure Lengend Snippet: A ChIP-qPCR analysis showing enrichment of E2F1 at the predicted response elements (RE1, RE2) in the PD-L1 promoter in MG63 and SW982 cells; enrichment is further increased upon E2F1 overexpression (Ac-H3, positive control; IgG, negative control). B , C Luciferase assays indicating that E2F1 enhances the activity of wild-type (WT) PD-L1 promoter constructs but not mutant (Mut) constructs lacking E2F1-binding sites. D , E Western blot and quantification of PDHA1, E2F1, and PD-L1 in sh-NC, sh-PDHA1, sh-PDHA1 + E2F1-OE, and E2F1-OE groups, showing that E2F1 rescues PD-L1 downregulation induced by PDHA1 knockdown. F , G Flow cytometry and statistical analysis of apoptosis demonstrating that PDHA1 knockdown increases apoptosis, which is partially reversed by E2F1 overexpression. H , I Western blot and quantification showing that PDHA1 knockdown and/or TTM treatment reduce Lip-DLAT, E2F1, and PD-L1 expression, with the strongest inhibition in the combination group. J , K Multiplex immunofluorescence and quantification in sarcoma tissues showing that high PDHA1 expression is associated with higher PD-L1 and reduced CD8⁺ T-cell infiltration. Scale bars: 50 μm. L Schematic model of the PDHA1–E2F1–PD-L1 axis in mediating CD8⁺ T-cell exclusion and immune evasion. Data are shown as mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: To modulate cuproptosis, MG63 cells with stable PDHA1 knockdown or control (sh-PDHA1 and sh-NC) were treated with the copper chelator ammonium tetrathiomolybdate (TTM; MedChemExpress, HY-100276).

Techniques: ChIP-qPCR, Over Expression, Positive Control, Negative Control, Luciferase, Activity Assay, Construct, Mutagenesis, Binding Assay, Western Blot, Knockdown, Flow Cytometry, Expressing, Inhibition, Multiplex Assay, Immunofluorescence

Journal: Cell metabolism

Article Title: Ejection of damaged mitochondria and their removal by macrophages ensure efficient thermogenesis in brown adipose tissue

doi: 10.1016/j.cmet.2022.02.016

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Pdhb ( {"type":"entrez-nucleotide","attrs":{"text":"NM_024221","term_id":"110227587","term_text":"NM_024221"}} NM_024221 ) Mouse Tagged ORF Clone Lentiviral Particle , OriGene Technologies , Cat# MR205484L3V.

Techniques: Immunofluorescence, Recombinant, Cytometry, Purification, Virus, Control, shRNA, Liposomes, Cell Culture, Red Blood Cell Lysis, XF Assay, Staining, Isolation, Transfection, Live Cell Imaging, Mouse Assay, Software, Gene Expression