human aml cell line u937  (ATCC)

Journal: Toxicology Reports

Article Title: Genotoxicity risk assessment of a 7-hydroxymitragynine-enriched Kratom preparation: An integrated in silico and in vitro approach

doi: 10.1016/j.toxrep.2026.102206

Figure Lengend Snippet: Micronucleus test of Kratom leaf extract after 4 h exposure with S9 in TK6 cells. Results are the mean ± SD of 3 independent experiments. Statistical testing with one-way ANOVA and Tukey’s post-hoc test (* p < 0.05).

Article Snippet: The human B lymphoblastoid cell line (TK6) (CRL-8015; batch No. 70045146), purchased from ATCC, was cultured in RPMI 1640 medium supplemented with 10 % fetal bovine serum (FBS) and 1 % penicillin/streptomycin.

Techniques:

Journal: Toxicology Reports

Article Title: Genotoxicity risk assessment of a 7-hydroxymitragynine-enriched Kratom preparation: An integrated in silico and in vitro approach

doi: 10.1016/j.toxrep.2026.102206

Figure Lengend Snippet: Micronucleus test of Kratom leaf extract after 4 h exposure without S9 in TK6 cells. Results are the mean ± SD of 3 independent experiments. Statistical testing with one-way ANOVA and Tukey’s post-hoc test (* p < 0.05).

Article Snippet: The human B lymphoblastoid cell line (TK6) (CRL-8015; batch No. 70045146), purchased from ATCC, was cultured in RPMI 1640 medium supplemented with 10 % fetal bovine serum (FBS) and 1 % penicillin/streptomycin.

Techniques:

Journal: Toxicology Reports

Article Title: Genotoxicity risk assessment of a 7-hydroxymitragynine-enriched Kratom preparation: An integrated in silico and in vitro approach

doi: 10.1016/j.toxrep.2026.102206

Figure Lengend Snippet: Micronucleus test of Kratom leaf extract after 24 h exposure without S9 in TK6 cells. Results are the mean ± SD of 3 independent experiments. Statistical testing with one-way ANOVA and Tukey’s post-hoc test (* p < 0.05).

Article Snippet: The human B lymphoblastoid cell line (TK6) (CRL-8015; batch No. 70045146), purchased from ATCC, was cultured in RPMI 1640 medium supplemented with 10 % fetal bovine serum (FBS) and 1 % penicillin/streptomycin.

Techniques:

Journal: Food Chemistry: Molecular Sciences

Article Title: Sangyod rice extract attenuates oleic acid–induced hepatic steatosis by modulating apoptotic, inflammatory, and lipid metabolic pathways

doi: 10.1016/j.fochms.2026.100387

Figure Lengend Snippet: Sangyod rice extract demonstrated a reduction in cytotoxicity and ROS levels in OA-induced HepG2 cells. (A) Viability of HepG2 cells exposed to different concentrations of Sangyod rice extract. (B) Viability of Sangyod rice extract treatment after OA-induced HepG2 cells. (C) ROS generation in OA-induced HepG2 cells. Results are presented as the mean ± SEM from four independent biological experiments ( n = 4). One-way ANOVA followed by Tukey ' s post hoc test was used to determine statistical significance. * p < 0.05 compared to the control group, and # p < 0.05 compared to the OA group. Groups: Control (0.1% DMSO); OA (0.4 mM), oleic acid-induced HepG2 cells without treatment; SR 10, OA-induced HepG2 cells +10 μg/mL Sangyod rice extract; SR 50, OA-induced HepG2 cells +50 μg/mL Sangyod rice extract; SR 100, OA-induced HepG2 cells +100 μg/mL Sangyod rice extract.

Article Snippet: The HepG2 human hepatocellular carcinoma cell line was procured from the American Type Culture Collection (Manassas, VA, USA) and nurtured in Dulbecco's modified Eagle's medium (Gibco, Waltham, MA, USA) enriched with 10% fetal bovine serum (Gibco, Waltham, MA, USA), 1% penicillin/streptomycin (Gibco, Waltham, MA, USA), and 1% l -glutamine (Gibco, Waltham, MA, USA).

Techniques: Control

Journal: Food Chemistry: Molecular Sciences

Article Title: Sangyod rice extract attenuates oleic acid–induced hepatic steatosis by modulating apoptotic, inflammatory, and lipid metabolic pathways

doi: 10.1016/j.fochms.2026.100387

Figure Lengend Snippet: Sangyod rice extract inhibited apoptosis in OA-induced HepG2 cells by suppressing the Bax and caspase-3 pathway. (A) Representative images of nuclei stained with Hoechst 33342. Images shown at ×20 magnification. Scale bar: 50 μm. (B) Percentage of apoptotic cells after treatment with Sangyod rice extract in OA-induced HepG2 cells. (C) Western blot analysis of Bax, Bcl-2, procaspase-3, and cleaved caspase-3. (D) Relative expression of Bax and Bcl-2. (E) Relative expression of procaspase 3, and cleaved caspase 3. Results are presented as the mean ± SEM from four independent biological experiments ( n = 4). One-way ANOVA followed by Tukey ' s post hoc test was used to determine statistical significance. *p < 0.05 compared to the control group, and #p < 0.05 compared to the OA group. Groups: Control (0.1% DMSO); OA (0.4 mM), oleic acid-induced HepG2 cells without treatment; SR 10, OA-induced HepG2 cells +10 μg/mL Sangyod rice extract; SR 50, OA-induced HepG2 cells +50 μg/mL Sangyod rice extract; SR 100, OA-induced HepG2 cells +100 μg/mL Sangyod rice extract.

Article Snippet: The HepG2 human hepatocellular carcinoma cell line was procured from the American Type Culture Collection (Manassas, VA, USA) and nurtured in Dulbecco's modified Eagle's medium (Gibco, Waltham, MA, USA) enriched with 10% fetal bovine serum (Gibco, Waltham, MA, USA), 1% penicillin/streptomycin (Gibco, Waltham, MA, USA), and 1% l -glutamine (Gibco, Waltham, MA, USA).

Techniques: Staining, Western Blot, Expressing, Control

Journal: Food Chemistry: Molecular Sciences

Article Title: Sangyod rice extract attenuates oleic acid–induced hepatic steatosis by modulating apoptotic, inflammatory, and lipid metabolic pathways

doi: 10.1016/j.fochms.2026.100387

Figure Lengend Snippet: Sangyod rice extract attenuated inflammation in OA-induced HepG2 cells through inhibition of the NF-κB pathway. (A) TNF-α gene, (B) IL-1β gene, (C) IL-6 gene, (D) IL-10 gene. (E) Western blot analysis of NF-κB. (F) Relative expression of NF-κB protein. Results are presented as the mean ± SEM from four independent biological experiments (n = 4). One-way ANOVA followed by Tukey ' s post hoc test was used to determine statistical significance. *p < 0.05 indicates significance compared to the control group, while #p < 0.05 denotes significance compared to the OA group. Groups: Control (0.1% DMSO); OA (0.4 mM), oleic acid-induced HepG2 cells without treatment; SR 10, OA-induced HepG2 cells +10 μg/mL Sangyod rice extract; SR 50, OA-induced HepG2 cells +50 μg/mL Sangyod rice extract; SR 100, OA-induced HepG2 cells +100 μg/mL Sangyod rice extract.

Article Snippet: The HepG2 human hepatocellular carcinoma cell line was procured from the American Type Culture Collection (Manassas, VA, USA) and nurtured in Dulbecco's modified Eagle's medium (Gibco, Waltham, MA, USA) enriched with 10% fetal bovine serum (Gibco, Waltham, MA, USA), 1% penicillin/streptomycin (Gibco, Waltham, MA, USA), and 1% l -glutamine (Gibco, Waltham, MA, USA).

Techniques: Inhibition, Western Blot, Expressing, Control

Journal: Food Chemistry: Molecular Sciences

Article Title: Sangyod rice extract attenuates oleic acid–induced hepatic steatosis by modulating apoptotic, inflammatory, and lipid metabolic pathways

doi: 10.1016/j.fochms.2026.100387

Figure Lengend Snippet: Sangyod rice extract reduced lipid accumulation in OA-induced HepG2 cells. (A) Oil Red O staining was conducted on HepG2 cells, with red fat droplets indicating lipid accumulation. Images shown at ×20 magnification. Scale bar: 50 μm. (B) Percentage of lipid accumulation post Oil Red O extraction. (C) Levels of TG were measured using an assay kit. The data is displayed as mean ± SEM from four independent biological experiments (n = 4). One-way ANOVA followed by Tukey ' s post hoc test was used to determine statistical significance. *p < 0.05 indicates significance compared to the control group, while #p < 0.05 denotes significance compared to the OA group. Groups: Control (0.1% DMSO); OA (0.4 mM), oleic acid-induced HepG2 cells without treatment; SR 10, OA-induced HepG2 cells +10 μg/mL Sangyod rice extract; SR 50, OA-induced HepG2 cells +50 μg/mL Sangyod rice extract; SR 100, OA-induced HepG2 cells +100 μg/mL Sangyod rice extract.

Article Snippet: The HepG2 human hepatocellular carcinoma cell line was procured from the American Type Culture Collection (Manassas, VA, USA) and nurtured in Dulbecco's modified Eagle's medium (Gibco, Waltham, MA, USA) enriched with 10% fetal bovine serum (Gibco, Waltham, MA, USA), 1% penicillin/streptomycin (Gibco, Waltham, MA, USA), and 1% l -glutamine (Gibco, Waltham, MA, USA).

Techniques: Staining, Extraction, Control

Journal: Food Chemistry: Molecular Sciences

Article Title: Sangyod rice extract attenuates oleic acid–induced hepatic steatosis by modulating apoptotic, inflammatory, and lipid metabolic pathways

doi: 10.1016/j.fochms.2026.100387

Figure Lengend Snippet: Effect of Sangyod rice extract on lipid metabolism in OA-induced HepG2 cells. (A) SREBP-1c gene (B) ACC gene, (C) FASN gene (D) CPT-1 A gene, (E) SCD1 gene, (F) MTTP gene. The data is displayed as mean ± SEM from four independent biological experiments (n = 4). One-way ANOVA followed by Tukey ' s post hoc test was used to determine statistical significance. *p < 0.05 indicates significance compared to the control group, while #p < 0.05 denotes significance compared to the OA group. Groups: Control (0.1% DMSO); OA (0.4 mM), oleic acid-induced HepG2 cells without treatment; SR 10, OA-induced HepG2 cells +10 μg/mL Sangyod rice extract; SR 50, OA-induced HepG2 cells +50 μg/mL Sangyod rice extract; SR 100, OA-induced HepG2 cells +100 μg/mL Sangyod rice extract.

Article Snippet: The HepG2 human hepatocellular carcinoma cell line was procured from the American Type Culture Collection (Manassas, VA, USA) and nurtured in Dulbecco's modified Eagle's medium (Gibco, Waltham, MA, USA) enriched with 10% fetal bovine serum (Gibco, Waltham, MA, USA), 1% penicillin/streptomycin (Gibco, Waltham, MA, USA), and 1% l -glutamine (Gibco, Waltham, MA, USA).

Techniques: Control

Journal: Food Chemistry: Molecular Sciences

Article Title: Sangyod rice extract attenuates oleic acid–induced hepatic steatosis by modulating apoptotic, inflammatory, and lipid metabolic pathways

doi: 10.1016/j.fochms.2026.100387

Figure Lengend Snippet: Effect of Sangyod rice extract on the expression of LPL-1, LPL-2, PGC-1α and PPARα in OA-induced HepG2 cells. (A) LPL-1 gene (B) LPL-2 gene, (C) PPARα gene (D) PGC-1α gene. The data is displayed as mean ± SEM from four independent biological experiments (n = 4). One-way ANOVA followed by Tukey ' s post hoc test was used to determine statistical significance. *p < 0.05 indicates significance compared to the control group, while #p < 0.05 denotes significance compared to the OA group. Groups: Control (0.1% DMSO); OA (0.4 mM), oleic acid-induced HepG2 cells without treatment; SR 10, OA-induced HepG2 cells +10 μg/mL Sangyod rice extract; SR 50, OA-induced HepG2 cells +50 μg/mL Sangyod rice extract; SR 100, OA-induced HepG2 cells +100 μg/mL Sangyod rice extract.

Article Snippet: The HepG2 human hepatocellular carcinoma cell line was procured from the American Type Culture Collection (Manassas, VA, USA) and nurtured in Dulbecco's modified Eagle's medium (Gibco, Waltham, MA, USA) enriched with 10% fetal bovine serum (Gibco, Waltham, MA, USA), 1% penicillin/streptomycin (Gibco, Waltham, MA, USA), and 1% l -glutamine (Gibco, Waltham, MA, USA).

Techniques: Expressing, Control

Journal: Food Chemistry: Molecular Sciences

Article Title: Sangyod rice extract attenuates oleic acid–induced hepatic steatosis by modulating apoptotic, inflammatory, and lipid metabolic pathways

doi: 10.1016/j.fochms.2026.100387

Figure Lengend Snippet: Sangyod rice extract regulates lipid metabolism through the Akt and MAPK signaling pathways. (A) Western blot analysis of Akt, ERK1/2 amd p38 MAPK, (B) Relative expression of pERK/ERK protein, (C) Relative expression of p-p38/p38 protein, (D) Relative expression of pAkt/Akt protein. The data is displayed as mean ± SEM from four independent biological experiments (n = 4). One-way ANOVA followed by Tukey ' s post hoc test was used to determine statistical significance. *p < 0.05 indicates significance compared to the control group, while #p < 0.05 denotes significance compared to the OA group. Groups: Control (0.1% DMSO); OA (0.4 mM), oleic acid-induced HepG2 cells without treatment; SR 10, OA-induced HepG2 cells +10 μg/mL Sangyod rice extract; SR 50, OA-induced HepG2 cells +50 μg/mL Sangyod rice extract; SR 100, OA-induced HepG2 cells +100 μg/mL Sangyod rice extract.

Article Snippet: The HepG2 human hepatocellular carcinoma cell line was procured from the American Type Culture Collection (Manassas, VA, USA) and nurtured in Dulbecco's modified Eagle's medium (Gibco, Waltham, MA, USA) enriched with 10% fetal bovine serum (Gibco, Waltham, MA, USA), 1% penicillin/streptomycin (Gibco, Waltham, MA, USA), and 1% l -glutamine (Gibco, Waltham, MA, USA).

Techniques: Protein-Protein interactions, Western Blot, Expressing, Control

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: International Journal of Molecular Medicine

Article Title: Ebastine targets HER2/HER3 signaling and cancer stem cell traits to overcome trastuzumab resistance in HER2-positive breast cancer

doi: 10.3892/ijmm.2026.5751

Figure Lengend Snippet: EBA impairs cancer stem cell-like properties. (A) BT474 and SKBR3 cells were treated with EBA for 48 h, and ALDH1 activity was assessed by flow cytometry using the Aldefluor assay. DEAB was used to define the baseline of Aldefluor-positive fluorescence. (B) BT474 cells (5x10 4 cells/ml) were plated in ultra-low attachment dishes and cultured in the presence or absence of EBA for 5 days. The number and volume of mammospheres were measured by microscopy. (C) Overall survival of patients with breast cancer stratified by the co-expression of ALDH1A1 and CD44. (D) Spearman correlation analysis of ALDH1A1 and CD44 mRNA levels in patients with HER2-positive breast cancer from The Cancer Genome Atlas cohort (n=76). Kaplan-Meier survival analyses of patients with HER2-overexpressing breast cancer stratified by (E) ALDH1A1 and (F) CD44 expression. Patients were divided into high- and low-expression groups based on the median gene expression. Statistical significance was determined using the log-rank test. (G) JIMT-1 cells were treated with EBA (3 μ M) for 48 h and the CD44 high /CD24 low cell populations were identified by flow cytometry. (H) JIMT-1 cells (1.5x10 4 cells/ml) were cultured under serum-free suspension conditions in the presence of EBA (3 μ M) for 8 days. Mammosphere number and volumes were quantified. ** P<0.01 and **** P<0.0001 vs. vehicle-treated control (0 μ M EBA). EBA, ebastine; ALDH, aldehyde dehydrogenase; DEAB, diethylaminobenzaldehyde; CTL, control; ISO, isotype.

Article Snippet: The human breast cancer cell lines SKBR3, BT474, MDA-MB-453 (American Type Culture Collection) and JIMT-1 (Leibnitz Institute DSMZ-German Collection of Microorganisms and Cell Cultures GmbH) were cultured in DMEM, MEM or RPMI-1640 (all Sigma-Aldrich; Merck KGaA) supplemented with 10% FBS (Gibco; Thermo Fisher Scientific, Inc.) and 100 U/ml penicillin-streptomycin at 37°C in a humidified atmosphere of 5% CO 2 .

Techniques: Activity Assay, Flow Cytometry, Fluorescence, Cell Culture, Microscopy, Expressing, Gene Expression, Suspension, Control

Journal: Molecular Metabolism

Article Title: A targeted metabolomic method to detect epigenetically relevant metabolites

doi: 10.1016/j.molmet.2026.102342

Figure Lengend Snippet: Distribution of acetyl-CoA isotopomers following [U–13C]glucose and [U–13C]glutamine incubation with an isogenic gain-of-function mutation (R132H) in the metabolic enzyme Isocitrate Dehydrogenase 1 (IDH1). U-87 MG human brain immortalized cell line, either wild-type or harboring the IDH1-R132H mutation, was incubated with [U– 13 C]glucose or [U– 13 C]glutamine for 20 and 72 h. The x axis represents the different acetyl-CoA isotopomers, showing the number of incorporated 13C atoms, with their origins within the acetyl-CoA structure noted in parentheses (see also ). The y axis indicates the percentage abundance of each isotopomer population, with n = 5 per condition after correction for natural abundance (refer to the Methods section 4.6.3). Statistical significance was assessed using Student’s t-test, with p -values represented as follows: ns: not significant ( p > 0.05); ∗: p ≤ 0.05. Because the M+2 labeling of the acetyl group may arise from either oxidative decarboxylation of pyruvate or reductive carboxylation of citrate, these two indistinguishable sources are jointly annotated as M+2 (Pyr/Cit). Abbreviations: Pyr/Cit, pyruvate- or citrate-derived M+2 (indistinguishable by MRM transitions); Gly, Glycine; For, Formate; R5P, Ribose-5-phosphate. # in x-axis indicates isotopomers with the same number of labeled carbon atoms incorporated, but with different origin and final moiety.

Article Snippet: The U-87 MG human brain immortalized cell line (ATCC® HTB-14TM) (wild type) or the R132H (IDH1 mutant) were plated at a density of 1 million cells per T75 plate in 10 mL complete ATCC Eagle’s Minimum Essential Medium (EMEM) 30-2003 medium, with 5 replicates per condition.

Techniques: Incubation, Mutagenesis, Labeling, Derivative Assay

Journal: Translational Oncology

Article Title: The VAX2-SERPINE1 axis modulates colorectal cancer cell proliferation and apoptosis through WNT/beta-catenin signaling

doi: 10.1016/j.tranon.2026.102703

Figure Lengend Snippet: VAX2 is upregulated and correlates with malignant progression in CRC. (A) Expression of VAX2 in the indicated CRC cell lines was determined by western blotting normalized to colon epithelial cell line FHC as a control. (B) Expression of VAX2 was determined in 13 pairs of CRC and normal tissues by western blotting using Wilcoxon signed-rank test. **, P < 0.05. (C) Expression of VAX2 was determined in 13 pairs of CRC and normal tissues by IHC. (D) Expression of VAX2 was determined in 79 pairs of CRC and normal tissues by IHC. (E) Quantitative expression of VAX2 in 79 pairs of CRC compared with normal tissues using Wilcoxon signed-rank test. ****, P < 0.001. (F) Quantitative expression of VAX2 in 99 CRC compared with 79 normal tissues using Mann–Whitney U test. ****, P < 0.001. (G) VAX2 expression levels in different subgroups stratified based on T stage. Mann–Whitney U test, **, P < 0.05. (H) VAX2 expression levels in different subgroups stratified based on AJCC stage. Mann–Whitney U test, **, P < 0.05. (I) Kaplan-Meier analyses of the overall survival probability in the TMA consisting of 99 patients based on VAX2 expression. Scale bars in C&D represent 100 μm.

Article Snippet: Normal colonic epithelial cell line FHC, human CRC cell lines (RKO, SW1116, HCT116 and SW620) and human GC cell lines (AGS) were purchased from the American Type Culture Collection (ATCC, USA; Catalog Numbers: FHC CRL-1831, RKO CRL-2577, SW1116 CCL-233, HCT116 CCL-247, SW620 CCL-227, AGS CRL-1739).

Techniques: Expressing, Western Blot, Control, MANN-WHITNEY

Journal: Translational Oncology

Article Title: The VAX2-SERPINE1 axis modulates colorectal cancer cell proliferation and apoptosis through WNT/beta-catenin signaling

doi: 10.1016/j.tranon.2026.102703

Figure Lengend Snippet: VAX2 facilitates CRC cell malignant behavior in vitro and in vivo. (A1/2) Expression of VAX2 in transfected LoVo, HCT116 and SW480 cell lines were analyzed by western blotting. (B1/2) Effects of VAX2 overexpression (B1) or knockdown (B2) on the proliferation of CRC cell lines, as determined by colony formation assay. ***, P < 0.01, Vector vs. VAX2; ***, P < 0.01, ****, P < 0.001, Scr-siRNA vs. VAX2-siRNAp. (C1/2) Representative graphs of EdU positivity in cells transfected with Vector and VAX2(C1) or Scr-siRNA and VAX2-siRNAp (C2). ***, P < 0.01, Vector vs. VAX2; ****, P < 0.001, Scr-siRNA vs. VAX2-siRNAp. (D1/2) Representative graphs of apoptosis assays in cells transfected with Vector and VAX2(D1) or Scr-siRNA and VAX2-siRNAp (D2). ****, P < 0.001, Vector vs. VAX2; ***, P < 0.01, ****, P < 0.001, Scr-siRNA vs. VAX2-siRNAp. (E1/2) Representative graphs of Transwell assays in cells transfected with Vector and VAX2(E1) or Scr-siRNA and VAX2-siRNAp (E2). ***, P < 0.01, Vector vs. VAX2; **, P < 0.05, ***, P < 0.01, Scr-siRNA vs. VAX2-siRNAp.

Article Snippet: Normal colonic epithelial cell line FHC, human CRC cell lines (RKO, SW1116, HCT116 and SW620) and human GC cell lines (AGS) were purchased from the American Type Culture Collection (ATCC, USA; Catalog Numbers: FHC CRL-1831, RKO CRL-2577, SW1116 CCL-233, HCT116 CCL-247, SW620 CCL-227, AGS CRL-1739).

Techniques: In Vitro, In Vivo, Expressing, Transfection, Western Blot, Over Expression, Knockdown, Colony Assay, Plasmid Preparation

Journal: Translational Oncology

Article Title: The VAX2-SERPINE1 axis modulates colorectal cancer cell proliferation and apoptosis through WNT/beta-catenin signaling

doi: 10.1016/j.tranon.2026.102703

Figure Lengend Snippet: VAX2 involves in WNT/beta-catenin signaling by promoting beta-catenin nucleus accumulation. (A&B) Kyoto Encyclopedia of Genes and Genomes (KEGG) (A) and Gene Set Enrichment Analysis (GSEA) (B) were conducted with differentially-expressed genes between Scr-siRNA and VAX2-siRNAp. (C) Western blotting of cytoplasmic and nuclear fractions of beta-catenin in VAX2 knockdown HCT116 cells. (D) Immunofluorescence of beta-catenin in VAX2 knockdown HCT116 and SW480 cells. (E) Co-immunoprecipitation of enrichment of beta-catenin binding to LEF1 in VAX2 knockdown HCT116 and SW480 cells. (F) qRT-PCR of abundance of beta-catenin-TCF/LEF-1 targets (MMP7, Cyclin D1 and c-jun) in VAX2 knockdown HCT116 and SW480 cells. **, P < 0.05, ***, P < 0.01, ****, P < 0.001, Scr-siRNA vs. VAX2-siRNAp. (G) VAX2-modulated CRC cells were transfected with the TOP/FOPflash reporter plasmid, and the reporter activities were determined 48 hours later ( n = 3). ***, P < 0.01, ****, P < 0.001, Scr-siRNA vs. VAX2-siRNAp. Yellow scale bar in D, 50 μm; green scale bar in D, 20 μm.

Article Snippet: Normal colonic epithelial cell line FHC, human CRC cell lines (RKO, SW1116, HCT116 and SW620) and human GC cell lines (AGS) were purchased from the American Type Culture Collection (ATCC, USA; Catalog Numbers: FHC CRL-1831, RKO CRL-2577, SW1116 CCL-233, HCT116 CCL-247, SW620 CCL-227, AGS CRL-1739).

Techniques: Western Blot, Knockdown, Immunofluorescence, Immunoprecipitation, Binding Assay, Quantitative RT-PCR, Transfection, Plasmid Preparation

Journal: Translational Oncology

Article Title: The VAX2-SERPINE1 axis modulates colorectal cancer cell proliferation and apoptosis through WNT/beta-catenin signaling

doi: 10.1016/j.tranon.2026.102703

Figure Lengend Snippet: VAX2 transcriptionally activates SERPINE1 in gastrointestinal cancers. (A) Heatmap of top 10 upregulated and downregulated genes between Scr-siRNA and VAX2-siRNAp. (B) Volcano plot of differentially-expressed genes between Scr-siRNA and VAX2-siRNAp. (C) SERPINE1 abundance was detected between Scr-siRNA and VAX2-siRNAp in CRC cell lines via qRT-PCR. ****, P < 0.001, Scr-siRNA vs. VAX2-siRNAp. (D) SERPINE1 abundance was detected between Vector and VAX2 in LoVo via qRT-PCR. ***, P < 0.01, Vector vs. VAX2. (E1/2) SERPINE1 abundance was detected between Vector and VAX2 in AGS (E1) and HGC-27 (E2) cells via qRT-PCR. **, P < 0.05, ****, P < 0.001, Vector vs. VAX2. (F) Enrichment site and binding motif of VAX2 on SERPINE1 promotor predicted by Jaspar database. (G) Chromatin immunoprecipitation (ChIP) was performed in CRC cell lines HCT116 and SW480 and GC cell lines AGS. (H) Wildtype (WT) or mutation (MUT) SERPINE1 promotor was cloned into luciferase plasmids. (I) The WT or MUT SERPINE1 promoter construct was co-transfected with VAX2-siRNAp or Scr-siRNA in HCT116 and SW480 cells, and the relative luciferase activity to corresponding control was determined ( n = 3). *, P >0.05, ***, P < 0.01, ****, P < 0.001, Scr-siRNA vs. VAX2-siRNAp. (J) The WT or MUT SERPINE1 promoter construct was co-transfected with VAX2 or Vector in AGS cells, and the relative luciferase activity to corresponding control was determined ( n = 3). *, P >0.05, ***, P < 0.01, ****, P < 0.001, Vector vs. VAX2.

Article Snippet: Normal colonic epithelial cell line FHC, human CRC cell lines (RKO, SW1116, HCT116 and SW620) and human GC cell lines (AGS) were purchased from the American Type Culture Collection (ATCC, USA; Catalog Numbers: FHC CRL-1831, RKO CRL-2577, SW1116 CCL-233, HCT116 CCL-247, SW620 CCL-227, AGS CRL-1739).

Techniques: Quantitative RT-PCR, Plasmid Preparation, Binding Assay, Chromatin Immunoprecipitation, Mutagenesis, Clone Assay, Luciferase, Construct, Transfection, Activity Assay, Control

Journal: Translational Oncology

Article Title: The VAX2-SERPINE1 axis modulates colorectal cancer cell proliferation and apoptosis through WNT/beta-catenin signaling

doi: 10.1016/j.tranon.2026.102703

Figure Lengend Snippet: VAX2 mediates CRC proliferation by transactivating SERPINE1 expression in vitro. (A) Proliferation of transfected CRC cell lines, as determined by colony formation assays. ****, P < 0.001. (B) Proliferation of transfected CRC cell lines, as determined by EdU assays. ****, P < 0.001. (C) Apoptosis of transfected CRC cell lines, as determined by apoptpsis assays using flow cytometry. (D) Co-immunoprecipitation of enrichment of beta-catenin binding to LEF1 in transfected HCT116 and SW480. (E) qRT-PCR of abundance of beta-catenin-TCF/LEF-1 targets (MMP7, Cyclin D1 and c-jun) in transfected HCT116 and SW480. **, P < 0.05, ***, P < 0.01, ****, P < 0.001.

Article Snippet: Normal colonic epithelial cell line FHC, human CRC cell lines (RKO, SW1116, HCT116 and SW620) and human GC cell lines (AGS) were purchased from the American Type Culture Collection (ATCC, USA; Catalog Numbers: FHC CRL-1831, RKO CRL-2577, SW1116 CCL-233, HCT116 CCL-247, SW620 CCL-227, AGS CRL-1739).

Techniques: Expressing, In Vitro, Transfection, Flow Cytometry, Immunoprecipitation, Binding Assay, Quantitative RT-PCR

Journal: Translational Oncology

Article Title: The VAX2-SERPINE1 axis modulates colorectal cancer cell proliferation and apoptosis through WNT/beta-catenin signaling

doi: 10.1016/j.tranon.2026.102703

Figure Lengend Snippet: VAX2 is correlated with SERPINE1 in human CRC tissues. (A) Expression of VAX2 was determined in 71 pairs of CRC and NT by qRT-PCR using waterfall plot. (B) Expression of SERPINE1 was determined in 71 pairs of CRC and NT by qRT-PCR using waterfall plot. (C) Expression of VAX2 was determined in 71 pairs of CRC and NT by qRT-PCR using box plot. ****, P < 0.001. (D) Expression of SERPINE1 was determined in 71 pairs of CRC and NT by qRT-PCR using box plot. **, P < 0.05. (E) Correlation between VAX2 and SERPINE1 in 71 pairs CRC tissues. (F) Correlation between VAX2 and SERPINE1 in TCGA-CRC. (G) VAX2 and SERPINE1 expression in CRC tissues were detected using HE and IHC. ( H) The signal transduction mechanism of VAX2. Scale bar in G, 100 μm.

Article Snippet: Normal colonic epithelial cell line FHC, human CRC cell lines (RKO, SW1116, HCT116 and SW620) and human GC cell lines (AGS) were purchased from the American Type Culture Collection (ATCC, USA; Catalog Numbers: FHC CRL-1831, RKO CRL-2577, SW1116 CCL-233, HCT116 CCL-247, SW620 CCL-227, AGS CRL-1739).

Techniques: Expressing, Quantitative RT-PCR, Transduction

Journal: The Journal of Biological Chemistry

Article Title: Dynamic assessment of the allocation of copper to cytochrome c oxidase using size-exclusion chromatography (SEC) combined with inductively coupled plasma mass spectrometry (ICP-MS)

doi: 10.1016/j.jbc.2026.111278

Figure Lengend Snippet: Identification and validation of CuCOX peak in SEC-ICP-MS chromatogram. A , Top : representative SEC-ICP-MS chromatograms of lysates from 786-O and RCC4 cells grown in 0.14 μM or 30 μM Cu for 48 h. Bottom: UV-Vis absorbance isoplots collected throughout the entire chromatography run. The legend shows the logarithmic scale for absorbance in the isoplot. Note that, because absorbance was measured before the ICP-MS detector, there is a slight shift in the timeline between the chromatogram and the spectra. B , measurement of absorbance from the 5 min fraction presented on a linear scale shows Cu-induced peaks at 420 and 600 nm. C , box whisker plots show relative abundance of Cu measured by SEC-ICP-MS for CuCOX and MTs, and by ICP-MS for total Cu content in 786-O (3 biological and 2 technical replicates) and RCC4 (4 biological and 2 technical replicates) cells. D , Western blot of indicated proteins eluted from the CuCOX peak and the inputs are shown below. E , oxygen consumption rate (OCR) from representative seahorse mitochondrial stress tests in 786-O and RCC4 cells (3 biological replicates). Quantification of basal and maximal respiration (post-FCCP injection), and respiration coupled to ATP production. Means ± SD are shown. p -values for basal respiration in 786-O cells was calculated using a two-tailed, paired t test. F , SEC-ICP-MS chromatograms, quantification of CuCOX and MT peaks, total Cu level, and Seahorse measurements of OCR for RPTEC TH1 and TERT1 cell lines (3 biological replicates and 2 technical replicates). Box-and-whisker plots display median, minimum and maximum values and all individual data points. Unless otherwise indicated, p -values were determined using a two-tailed, unpaired t test. All Cu measurements are normalized to total phosphorus (P). HMW, high molecular weight fraction; LMW, low molecular weight fraction; MT, metallothioneins

Article Snippet: Human renal carcinoma cell lines 786-O (RRID: CVCL_1051, ATCC) and RCC4 (RRID: CVCL_0498) and human immortalized renal proximal tubule cells RPTEC cells TH1 (kerafast ECH001) (RRID: CVCL_K278) and TERT1 (ATCC CRL-4031were cultured in DMEM/F12 medium (Cytiva, SH30023) supplemented with 10% fetal bovine serum (FBS; Gibco, 6000–044) at 37 °C in a humidified atmosphere containing 5% CO 2 .

Techniques: Biomarker Discovery, Chromatography, Whisker Assay, Western Blot, Injection, Two Tailed Test, High Molecular Weight, Molecular Weight

Journal: The Journal of Biological Chemistry

Article Title: Dynamic assessment of the allocation of copper to cytochrome c oxidase using size-exclusion chromatography (SEC) combined with inductively coupled plasma mass spectrometry (ICP-MS)

doi: 10.1016/j.jbc.2026.111278

Figure Lengend Snippet: Galactose induces oxidative phosphorylation and increases Cu content and allocation to CuCOX in 786-O RCC cells. A , schematic representation of the Leloir pathway converting galactose into glucose. B , OCR and mitochondrial ATP production in response to 48 h treatment with galactose ( red ) as compared to glucose ( black ) (3 biological replicates). C , SEC-ICP-MS chromatograms of Cu ( top ) and corresponding UV-Vis absorbance plots in lysates from 786-O cells treated with 10 mM glucose or 10 mM galactose for 48 h. The legend shows the logarithmic scale for absorbance in the isoplot. D , box whiskers show quantification of Cu allocated to CuCOX and MTs, and total Cu content in lysates from 786-O cells grown in glucose- or galactose-containing media (7 biological replicates with 2 technical replicates for each). E , Western blot of total input lysates and CuCOX eluates from 786-O grown in glucose or galactose media. F , Schematic model of mitochondrial electron transport chain along with localization of inhibitors for each complex. G , quantification of SEC-ICP-MS peaks corresponding to Cu in CuCOX and MTs, and total Cu in lysates of 786-O cells treated with media containing glucose, galactose, or galactose with the indicated electron transport chain inhibitors added for 3 h before collection. ROT, rotenone (200 nM, three biological replicates in technical duplicates), IACS, IACS-10759 (1 nM, 3biological replicates in technical duplicates), MET, metformin (25 mM, 3 biological replicates in technical duplicates), AM, antimycin A (1 nM, 4 biological replicates in technical duplicates), MYX, myxothiazol (1 nM, 4 biological replicates run in technical duplicates), KCN, potassium cyanide (0.5 mM, 4 biological replicates in technical duplicates), NaN3, sodium azide (0.5 mM, 4 biological replicates in technical duplicates). Box-and-whisker plots displayed the median, minimum and maximum values, and all individual data points. All p -values were calculated using a two-tailed, unpaired t test and compared the treatment groups with the galactose-only condition.

Article Snippet: Human renal carcinoma cell lines 786-O (RRID: CVCL_1051, ATCC) and RCC4 (RRID: CVCL_0498) and human immortalized renal proximal tubule cells RPTEC cells TH1 (kerafast ECH001) (RRID: CVCL_K278) and TERT1 (ATCC CRL-4031were cultured in DMEM/F12 medium (Cytiva, SH30023) supplemented with 10% fetal bovine serum (FBS; Gibco, 6000–044) at 37 °C in a humidified atmosphere containing 5% CO 2 .

Techniques: Phospho-proteomics, Western Blot, Whisker Assay, Two Tailed Test

Journal: The Journal of Biological Chemistry

Article Title: Dynamic assessment of the allocation of copper to cytochrome c oxidase using size-exclusion chromatography (SEC) combined with inductively coupled plasma mass spectrometry (ICP-MS)

doi: 10.1016/j.jbc.2026.111278

Figure Lengend Snippet: Kinetics of allocation of 63Cu tracer into CuCOX. A , schematic representation of tracing 63 Cu. B , timeline of 63 Cu tracking into CuCOX. C and D , SEC-ICP-MS chromatograms of exogenous 63 Cu in lysates from 786-O and RCC4 cells at the indicated time points. E and F , quantification by SEC-ICP-MS of exogenous 63 Cu in CuCOX, MTs and total exogenous 63 Cu in 786-O and RCC4 cells. Cu abundance was normalized to Phosphorus (P). In all experiments: n = 4, SEM ± SD are shown; p- values were calculated using a two-tailed, unpaired t test and indicate significance of the difference between 10 or 30 μM Cu compared to 0.14 μM Cu. Experiments were performed in 3 technical replicates twice and a representative experiment is shown.

Article Snippet: Human renal carcinoma cell lines 786-O (RRID: CVCL_1051, ATCC) and RCC4 (RRID: CVCL_0498) and human immortalized renal proximal tubule cells RPTEC cells TH1 (kerafast ECH001) (RRID: CVCL_K278) and TERT1 (ATCC CRL-4031were cultured in DMEM/F12 medium (Cytiva, SH30023) supplemented with 10% fetal bovine serum (FBS; Gibco, 6000–044) at 37 °C in a humidified atmosphere containing 5% CO 2 .

Techniques: Two Tailed Test

Journal: The Journal of Biological Chemistry

Article Title: Dynamic assessment of the allocation of copper to cytochrome c oxidase using size-exclusion chromatography (SEC) combined with inductively coupled plasma mass spectrometry (ICP-MS)

doi: 10.1016/j.jbc.2026.111278

Figure Lengend Snippet: Allocation of tracer 63 Cu to CuCOX in 786-O and RCC4 cells chronically grown in low or high Cu media. A , timeline of the experiment: AC, exposure to 30 μM 63 Cu for 48 h of cells grown continuously in low Cu media; CH, exposure to 30 μM 63 Cu for 48 h of cells grown in high Cu media. B , representative examples of Cu SEC-ICP-MS chromatograms from cells grown in Cu low media and exposed to 30 μM 63 Cu for 48 h (AC) or cells grown continuously in media containing 30 μM Cu and then treated with the same concentration of 63 Cu (CH). C , quantification of 63 Cu in total exogenous and endogenous Cu measured by SEC-ICP-MS. D , quantification of 63 Cu in exogenous and endogenous CuCOX peak from SEC-ICP-MS chromatograms. E , representative examples of Zn SEC-ICP-MS chromatograms as described in B. F , quantification of total cellular Zn based on SEC-ICP-MS. G , representative examples of Fe SEC-ICP-MS chromatograms as described in B . H , quantification of total cellular Fe based on SEC-ICP-MS chromatograms. Metals’ abundance was normalized to Phosphorus (P). In all experiments: Means ± SD are shown; n = 4 biological replicates; p - values were calculated using a two-tailed, unpaired t test.

Article Snippet: Human renal carcinoma cell lines 786-O (RRID: CVCL_1051, ATCC) and RCC4 (RRID: CVCL_0498) and human immortalized renal proximal tubule cells RPTEC cells TH1 (kerafast ECH001) (RRID: CVCL_K278) and TERT1 (ATCC CRL-4031were cultured in DMEM/F12 medium (Cytiva, SH30023) supplemented with 10% fetal bovine serum (FBS; Gibco, 6000–044) at 37 °C in a humidified atmosphere containing 5% CO 2 .

Techniques: Concentration Assay, Two Tailed Test

Journal: The Journal of Biological Chemistry

Article Title: Dynamic assessment of the allocation of copper to cytochrome c oxidase using size-exclusion chromatography (SEC) combined with inductively coupled plasma mass spectrometry (ICP-MS)

doi: 10.1016/j.jbc.2026.111278

Figure Lengend Snippet: Contribution of Cu transporters to the allocation of tracer 63 Cu to CuCOX. A , timeline of experiments for cells cultured under low or high Cu conditions and subsequently exposed to 30 μM 63 Cu for 48 h (AC and CH, respectively) or to 0.14 μM 63 Cu for 48 h (LO). B–E , effects of the knockdowns of indicated transporters on the allocation of exogenous 63 Cu and total content of Zn and Fe measured by SEC-ICP-MS in AC conditions for 786-O ( top ) and RCC4 ( bottom ) cells. F-I , effects of the knockdowns of indicated transporters on the allocation of exogenous 63 Cu and total content of Zn and Fe measured by SEC-ICP-MS in CH conditions for both cell lines. J and K , Western blots showing efficiency of the indicated knockdowns for AC ( J ) and CH ( K ) conditions. L and M , RT-PCR validation of CTR1 KD under AC ( L ) and CH ( M ) conditions. NT-non-targeting siRNA. Metals’ abundance was normalized to Phosphorus (P). N , Western blots showing expression of ZNT1 in RCC4 and 786-O cells and the effectiveness of the knockout. O , effects of the ZNT1 knockout on the total exogenous 63 Cu uptake and 63 Cu allocation to CuCOX, as well as total content of Zn, and Fe measured by SEC-ICP-MS in AC conditions for RCC4 cells. P , SEC-ICP-MS chromatograms of lysates from 786-O cells grown in 0.14 μM Cu, treated with non-targeting (NT) or CTR1 siRNAs, and exposed to 0.14 μM 63 Cu for 48 h. Q , quantification of total 63 Cu, and 63 Cu allocated to CuCOX and MT measured by SEC-ICP-MS in response to CTR1 KD. R , DepMap gene-effect scores distribution and median across all RCC cell lines for five metal transporter genes. A blue dot marks 786-O cell line. Biological replicates: B–I and Q , n = 4 (except for NT treatment in RCC4 in B–E and CTR1 KD in 786-O in F–I where n = 3); L and M , n = 3; O, n = 6. Means ± SD are shown; p - values were calculated using a two-tailed, unpaired t test and comparing the significance of changes caused by specific si/sgRNA to NT si/sgRNA.

Article Snippet: Human renal carcinoma cell lines 786-O (RRID: CVCL_1051, ATCC) and RCC4 (RRID: CVCL_0498) and human immortalized renal proximal tubule cells RPTEC cells TH1 (kerafast ECH001) (RRID: CVCL_K278) and TERT1 (ATCC CRL-4031were cultured in DMEM/F12 medium (Cytiva, SH30023) supplemented with 10% fetal bovine serum (FBS; Gibco, 6000–044) at 37 °C in a humidified atmosphere containing 5% CO 2 .

Techniques: Cell Culture, Western Blot, Reverse Transcription Polymerase Chain Reaction, Biomarker Discovery, Expressing, Knock-Out, Two Tailed Test