human osteosarcoma u2os  (ATCC)

Journal: Genes & Diseases

Article Title: Network pharmacology reveals that Yanghe Decoction inhibits osteosarcoma progression via ROS-induced mitochondrial dysfunction and enhances cisplatin sensitivity

doi: 10.1016/j.gendis.2025.101862

Figure Lengend Snippet: Identification of active compounds and target prediction in YHD. (A) Venn diagram of the target of YHD and the target of osteosarcoma. (B – D) Gene Ontology (GO) enrichment analysis results. (E, F) Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis results. (G) The component-target-pathway-disease network implicated in the mechanism of YHD in osteosarcoma treatment. The triangles represent osteosarcoma, the diamonds represent pathways, the circles represent key genes, and the squares represent the active ingredients of YHD. (H) Heatmap of molecular docking score. A binding energy heatmap with a bluer color indicates a more stable binding. (I) Molecular docking visualization between the active components of YHD and key targets.

Article Snippet: Human osteosarcoma cell lines (HOS, 143B), human bone marrow stromal cells (HS-5), human proximal renal tubular epithelial cells (HK-2), and human normal liver cells (LO2) were all purchased from the American Type Culture Collection (ATCC).

Techniques: Binding Assay

Journal: Genes & Diseases

Article Title: Network pharmacology reveals that Yanghe Decoction inhibits osteosarcoma progression via ROS-induced mitochondrial dysfunction and enhances cisplatin sensitivity

doi: 10.1016/j.gendis.2025.101862

Figure Lengend Snippet: YHD selectively inhibits osteosarcoma (OS) cells without affecting the viability or apoptosis of normal human cells. (A, B) CCK8 assay detected the effect of YHD on the viability of OS cells at 24 h and 48 h. (C, D) Colony formation assay detected the effect of YHD on the colony-forming ability of OS cells. (E – G) Flow cytometry was used to detect the effect of YHD on the cell cycle of OS cells. (H – J) Western blot analysis detected the effect of YHD on the levels of proliferation-related proteins in OS cells. Data were presented as mean ± standard deviation ( n = 3). ∗ p < 0.05 and ∗∗ p < 0.01 versus the blank group.

Article Snippet: Human osteosarcoma cell lines (HOS, 143B), human bone marrow stromal cells (HS-5), human proximal renal tubular epithelial cells (HK-2), and human normal liver cells (LO2) were all purchased from the American Type Culture Collection (ATCC).

Techniques: CCK-8 Assay, Colony Assay, Flow Cytometry, Western Blot, Standard Deviation

Journal: Genes & Diseases

Article Title: Network pharmacology reveals that Yanghe Decoction inhibits osteosarcoma progression via ROS-induced mitochondrial dysfunction and enhances cisplatin sensitivity

doi: 10.1016/j.gendis.2025.101862

Figure Lengend Snippet: YHD can inhibit the migration and invasion of osteosarcoma (OS) cells, and promote their apoptosis. (A, B) Scratch healing assay showed that YHD inhibited the migration of OS cells. (C, D) Transwell assay showed that YHD inhibited the invasion of OS cells. (E – G) Western blot analysis detected the effect of YHD on the levels of proteins related to migration and invasion in OS cells. (H–K) Flow cytometry was used to detect the effect of YHD on apoptosis in OS cells. (L – N) Western blot analysis detected the effect of YHD on the levels of apoptosis-related proteins in OS cells. Data were presented as mean ± standard deviation ( n = 3). ∗ p < 0.05 and ∗∗ p < 0.01 versus the blank group.

Article Snippet: Human osteosarcoma cell lines (HOS, 143B), human bone marrow stromal cells (HS-5), human proximal renal tubular epithelial cells (HK-2), and human normal liver cells (LO2) were all purchased from the American Type Culture Collection (ATCC).

Techniques: Migration, Transwell Assay, Western Blot, Flow Cytometry, Standard Deviation

Journal: Genes & Diseases

Article Title: Network pharmacology reveals that Yanghe Decoction inhibits osteosarcoma progression via ROS-induced mitochondrial dysfunction and enhances cisplatin sensitivity

doi: 10.1016/j.gendis.2025.101862

Figure Lengend Snippet: YHD induces osteosarcoma (OS) cell death by increasing ROS levels. (A) The effect of YHD on ROS levels in OS cells was detected using the DCFH probe method. (B) After N-acetylcysteine (NAC) treatment, the effect of YHD on ROS levels in OS cells was detected using the DCFH probe method. (C) After NAC treatment, CCK8 assay detected the effect of YHD on the viability of OS cells at 24 h and 48 h. (D, E) After NAC treatment, scratch healing assay showed that YHD inhibited the migration of OS cells. (F, G) After NAC treatment, Transwell assay showed that YHD inhibited the invasion of OS cells. (H, I) After NAC treatment, flow cytometry was used to detect the effect of YHD on the apoptosis of OS cells. Data were presented as mean ± standard deviation ( n = 3). ∗ p < 0.05 and ∗∗ p < 0.01 versus the blank group.

Article Snippet: Human osteosarcoma cell lines (HOS, 143B), human bone marrow stromal cells (HS-5), human proximal renal tubular epithelial cells (HK-2), and human normal liver cells (LO2) were all purchased from the American Type Culture Collection (ATCC).

Techniques: CCK-8 Assay, Migration, Transwell Assay, Flow Cytometry, Standard Deviation

Journal: Genes & Diseases

Article Title: Network pharmacology reveals that Yanghe Decoction inhibits osteosarcoma progression via ROS-induced mitochondrial dysfunction and enhances cisplatin sensitivity

doi: 10.1016/j.gendis.2025.101862

Figure Lengend Snippet: YHD can induce mitochondrial dysfunction in osteosarcoma (OS) cells. (A) Real-time quantitative PCR was used to measure mitochondrial DNA (mtDNA) levels. (B, C) Western blot analysis detected the effect of YHD on the expression levels of proteins related to mitochondrial biogenesis in OS cells. (D, E) JC-1 staining detected the effect of YHD on the mitochondrial membrane potential in OS cells. (F, G) MitoSOX staining detected the effect of YHD on mitochondrial ROS levels in OS cells. (H) Seahorse XFe24 analyzer measured the effect of YHD on the oxygen consumption rate (OCR) in OS cells. (I) The effect of YHD on ATP content in OS cells. Data were presented as mean ± standard deviation ( n = 3). ∗ p < 0.05 and ∗∗ p < 0.01 versus the blank group.

Article Snippet: Human osteosarcoma cell lines (HOS, 143B), human bone marrow stromal cells (HS-5), human proximal renal tubular epithelial cells (HK-2), and human normal liver cells (LO2) were all purchased from the American Type Culture Collection (ATCC).

Techniques: Real-time Polymerase Chain Reaction, Western Blot, Expressing, Staining, Membrane, Standard Deviation

Journal: Genes & Diseases

Article Title: Network pharmacology reveals that Yanghe Decoction inhibits osteosarcoma progression via ROS-induced mitochondrial dysfunction and enhances cisplatin sensitivity

doi: 10.1016/j.gendis.2025.101862

Figure Lengend Snippet: YHD exerts anti-tumor effects on osteosarcoma (OS) cells through the PI3K/AKT and p38 signaling pathways. (A) Principal component analysis revealed a clear distinction in gene expression profiles between the control and YHD groups. (B) Volcano plot identified 3495 differentially expressed genes in the YHD group. (C) Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses. (D – G) Gene Set Enrichment Analysis (GSEA) of control and YHD groups. (H, I) Western blot analysis detected the effect of YHD on proteins related to the PI3K/AKT and MAPK pathways in OS cells. (J, K) After the addition of a PI3K activator and a P38 inhibitor, scratch healing assay showed that YHD inhibited the migration of OS cells. (L, M) After the addition of a PI3K activator and a P38 inhibitor, JC-1 staining detected the effect of YHD on the mitochondrial membrane potential in OS cells. Data were presented as mean ± standard deviation ( n = 3). ∗ p < 0.05 and ∗∗ p < 0.01 versus the blank group.

Article Snippet: Human osteosarcoma cell lines (HOS, 143B), human bone marrow stromal cells (HS-5), human proximal renal tubular epithelial cells (HK-2), and human normal liver cells (LO2) were all purchased from the American Type Culture Collection (ATCC).

Techniques: Protein-Protein interactions, Gene Expression, Control, Western Blot, Migration, Staining, Membrane, Standard Deviation

Journal: bioRxiv

Article Title: Proximity labelling of the BAK macropore uncovers a new role for SLC35A4-MP in mitochondrial dynamics

doi: 10.64898/2026.03.22.713508

Figure Lengend Snippet: A . SDS-PAGE separation and immunoblotting of mitochondria isolated from WT, SLC35A4–MP KO and SLC35A4-MP KO + SLC35A4-MP FLAG U2OS cells. SLC35A4-MP FLAG expression was induced with 5 ng/ml doxycycline for 48 h. SDHA serves as a loading control. Representative of n = 3 biological replicates. B. Representative confocal images of SLC35A4-MP KO + SLC35A4-MP FLAG U2OS cells. SLC35A4-MP FLAG expression was induced with 5 ng/ml doxycycline for 48 h. Immunofluorescence was used to label mitochondria (Anti-TOM20, magenta), and determine SLC35A4-MP localisation (anti-FLAG, yellow). Scale bar indicates 5 µm. C. Electron microscopy of WT and SLC35A4-MP KO U2OS cells. Scale bars indicate 500 nm. D. The steady-state whole cell proteome of SLC35A4-MP KO vs WT. MitoCarta 3.0 proteins are highlighted in black, and those with OMM or IMM localisations are highlighted in blue and pink respectively. E. Whole cell proteome of SLC35A4-MP KO vs WT U2OS in apoptotic vs non-apoptotic conditions. The log 2 (fold change) of significantly altered ( p < 0.05) WT and KO proteins following 1 h apoptotic treatment was plotted against each other; WT (x-axis) and KO (y-axis). Mitochondrial proteins are highlighted in black, and those with OMM or IMS/IMM localisation are highlighted in blue or pink respectively.

Article Snippet: Human osteosarcoma cells (U2OS) were purchased from the ATCC, and a clonal line was established.

Techniques: SDS Page, Western Blot, Isolation, Expressing, Control, Immunofluorescence, Electron Microscopy

Journal: bioRxiv

Article Title: Proximity labelling of the BAK macropore uncovers a new role for SLC35A4-MP in mitochondrial dynamics

doi: 10.64898/2026.03.22.713508

Figure Lengend Snippet: A . The steady-state SLC35A4-MP interactome. Proteins with a log 2 (SLC35A4-MP FLAG 0 h / WT 0 h) > 1, p value < 0.05, were considered significantly enriched. Proteins with MitoCarta 3.0 annotations have been coloured as indicated B. Comparison of log 2 (SLC35A4-MP FLAG / WT) enrichment of the SLC35A4-MP FLAG interactome at 0 h and 1 h apoptosis. Mitochondrial proteins are coloured in black. Proteins preferentially enriched at 0 h or 1 h post induction of apoptosis are highlighted by the blue and red regions respectively. Stable interactors are highlighted by the yellow region (refer to Fig. S4B) C-D. BN-PAGE analysis of 1% digitonin soluble complexes from isolated mitochondria, immunoblotting using antibodies against C) MIC10 (MICOS complex) and D) OPA1 respectively. NDUFA9 (CI) and SDHA (CII) serve as a loading controls. Representative of n = 3 independent experiments. For densitometric quantification of soluble MIC10 or OPA1 containing complexes, complex abundance was normalised to NDUFA9 or SDHA respectively and displayed relative to WT levels. Data represents n = 3 independent experiments. Error bars indicate mean ± SEM. * = p value < 0.05, ** = p value < 0.01 by one-way ANOVA with Tukey’s multiple comparison test. E. SDS-PAGE analysis of OPA1 isoforms in mitochondria isolated from control, SLC35A4-MP KO and SLC35A4-MP rescue U2OS cells (SLC35A4-MP FLAG) . SLC35a4-MP FLAG expression was induced with doxycycline for 48 h (concentrations as indicated). LONP1 serves as a loading control. Representative of n = 3 independent experiments. F-G. Densitometric quantification of L-OPA1 ‘a’ isoform, and S-OPA1 isoforms ‘c’ and ‘e’ in F) SLC35A4-MP KO (without doxycycline treatment), or G) SLC35A4-MP KO cells overexpressing SLC35A4-MP FLAG with increasing concentrations of doxycycline. OPA1 isoform levels were normalised to LONP1 and displayed relative to WT (without doxycycline treatment). n = 3 independent experiments. Error bars indicate mean ± SEM. F: *** = p value < 0.005 by Student’s t-test. G: ** = p value < 0.01 *** = p value < 0.005 by one-way ANOVA with Tukey’s multiple comparisons test.

Article Snippet: Human osteosarcoma cells (U2OS) were purchased from the ATCC, and a clonal line was established.

Techniques: Comparison, Isolation, Western Blot, SDS Page, Control, Expressing

Journal: bioRxiv

Article Title: Proximity labelling of the BAK macropore uncovers a new role for SLC35A4-MP in mitochondrial dynamics

doi: 10.64898/2026.03.22.713508

Figure Lengend Snippet: A. Live-cell imaging of WT, SLC35A4-MP KO and SLC35A4-MP rescue U2OS cells (SLC35A4-MP FLAG) stably expressing TOM20 Halo following treatment with ABT-737 [10 µM], S63845 [2 µM] and 1 h pre-treatment with QVD-OPh [20 µM]. Representative images from n=3 independent experiments. Scale bars indicate 5 µm. B-C. Quantification of apoptosis induced mtDNA release and mitochondrial fragmentation respectively. Quantification was performed on 10-15 cells from n = 3 independent experiments. Data points indicate mean ± SEM. Dashed line indicates the start of image acquisition D. Live cell imaging of SLC35A4-MP KO U2OS cells following CCCP [20µM] treatment. Representative images from n = 3 independent experiments. Scale bars indicate 10 µm. E. Quantification of mitochondrial fragmentation. Quantification was performed on 10-15 cells from n=3 independent experiments. Dashed line indicates start of image acquisition.

Article Snippet: Human osteosarcoma cells (U2OS) were purchased from the ATCC, and a clonal line was established.

Techniques: Live Cell Imaging, Stable Transfection, Expressing

Journal: iScience

Article Title: Predicting DNA damage response using synthetic cell painting profiles and experimental analysis

doi: 10.1016/j.isci.2026.115000

Figure Lengend Snippet: Application of the model to external datasets and experimental testing (A) Principal component analysis plot for the idr-0080 and cpg-0012 datasets. (B) Classification of cpg-0012 compounds into high-DDR (9,923) and low-DDR (20,694) groups. The high-DDR group includes known DNA damage inducers. Test compounds without prior DDR annotation were selected from the high-DDR (tetrindole, KF38789, and LY2183240) and low-DDR (amoxapine, acetazolamide, and captopril) groups. Flow cytometric (C) and Western blot (D) analyses of γH2AX expression in U2OS cells following 18 h treatment with 10 μM test compounds. (E) U2OS cells were treated with high-DDR candidates at increasing doses or with 10 μM low-DDR candidates for 72 h, and cell viability was measured using the CellTiter-Glo assay. Data are shown as means ± SD from independent experiments. One-way ANOVA with LSD post hoc test was used for statistical analysis. ∗ , p < 0.05; ∗∗∗ , p < 0.001 versus vehicle.

Article Snippet: U2OS human osteosarcoma cells (ATCC, HTB-96), originally derived from a moderately differentiated sarcoma of the tibia of a 15-year-old White female osteosarcoma patient, were obtained from the American Type Culture Collection (ATCC; Manassas, VA, USA) and authenticated by ATCC-provided STR profiling.

Techniques: Western Blot, Expressing, Glo Assay

Journal: iScience

Article Title: Predicting DNA damage response using synthetic cell painting profiles and experimental analysis

doi: 10.1016/j.isci.2026.115000

Figure Lengend Snippet:

Article Snippet: U2OS human osteosarcoma cells (ATCC, HTB-96), originally derived from a moderately differentiated sarcoma of the tibia of a 15-year-old White female osteosarcoma patient, were obtained from the American Type Culture Collection (ATCC; Manassas, VA, USA) and authenticated by ATCC-provided STR profiling.

Techniques: Recombinant, Flow Cytometry, Software