cuso 4  (Labeo Technologies Inc)

Journal: Molecular and Cellular Biochemistry

Article Title: High copper levels induce oxidative stress and inflammatory processes in a cell culture model of Wilson’s disease

doi: 10.1007/s11010-026-05481-6

Figure Lengend Snippet: Multivariate analysis of RNA sequencing data. The GSE107323 dataset includes HepG2 and HepG2 ATP7B-KO cells that were treated with 0.5mmol/l CuCl 2 for 6–24 h. Mean gene counts > 30 were used for analysis. a Principal component analysis was performed with the following settings for standard deviation: p = 0.005 and q = 0,01788 (FDR). b Thus, 2,142 genes were hierarchically clustered via a multivariance analysis visualised in a heatmap. c Genes were analysed by functional annotation analysis for biological processes by the DAVID tool and showed high EASE scores in genes involved in autophagy ( p = 1.1E-7), lipid metabolism ( p = 3.0E-6) and stress response ( p = 8,6E-5). Volcano plots were generated to compare the CuCl 2 -dependent gene regulation, indicating d 7,722 gene alterations in HepG2 cells and e 7,241 gene alterations in HepG2 ATP7B-KO cells. sig, significant; ut, untreated; t, treated

Article Snippet: Cell viability of CuCl 2 (Merck) or CuSO 4 (Merck) -treated HepG2 and HepG2 ATP7B-KO cells was measurement by cell counting assay (CCK-8, Sigma-Aldrich).

Techniques: RNA Sequencing, Standard Deviation, Functional Assay, Generated

Journal: Molecular and Cellular Biochemistry

Article Title: High copper levels induce oxidative stress and inflammatory processes in a cell culture model of Wilson’s disease

doi: 10.1007/s11010-026-05481-6

Figure Lengend Snippet: CuCl 2 -dependent gene expression of autophagy-associated genes. HepG2 and HepG2 ATP7B-KO cells were treated with 0.2–1.2.2mmol/l CuCl 2 or CuSO 4 for 6–24 h. Cell viability was measured after a CuCl 2 and b CuSO 4 treatment via a CCK-8 assay and normalised to that of the untreated control (mean ± SD; n = 4 (CuCl 2 ), n = 5 (CuSO 4 )). RNA from HepG2 and HepG2 ATP7B-KO cells treated with CuCl 2 and untreated PHH (Ctrl and WD) was extracted, and the expression of autophagy-related genes ( c , GABARALP1 ; d , HSP90AA1 ; e , LAMTOR3 ; f , PLOD2 ; g , PRAGC ; h , UBC ) was measured via two-step qRT‒PCR (mean ± SD; n = 3 (HepG2 and HepG2 ATP7B-KO), n = 4 (PHH Ctrl), n = 2 (PHH WD). The data represent copy numbers normalised to 100,000 copies of the reference gene GAPDH . The effect of CuCl 2 concentrations in HepG2 or HepG2 ATP7B-KO cells (* p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001) and differences between HepG2 and HepG2 ATP7B-KO cells after 6–24 h (# p < 0.05; ## p < 0.01; ### p < 0.001; ####; p < 0.0001) were measured by two-way ANOVA tests and Tukey’s multiple comparison tests. Differences in gene expression between PHH Ctrl and WD cells were tested by an unpaired t-test (× p < 0.05; ×× p < 0.01). Ctrl control, PHH primary human hepatocytes, WD Wilson’s disease

Article Snippet: Cell viability of CuCl 2 (Merck) or CuSO 4 (Merck) -treated HepG2 and HepG2 ATP7B-KO cells was measurement by cell counting assay (CCK-8, Sigma-Aldrich).

Techniques: Gene Expression, CCK-8 Assay, Control, Expressing, Comparison

Journal: Molecular and Cellular Biochemistry

Article Title: High copper levels induce oxidative stress and inflammatory processes in a cell culture model of Wilson’s disease

doi: 10.1007/s11010-026-05481-6

Figure Lengend Snippet: Copper induces oxidative stress and H 2 O 2 production. HepG2 and HepG2 ATP7B-KO cells were treated with 0.2–0.6mmol/l CuCl 2 for 6–24 h. RNA from HepG2 and HepG2 ATP7B-KO cells and PHH (Ctrl and WD) was extracted, and the gene expression of oxidated stress-related genes ( a , HMOX1 ; b , DNAJB1 ; c , GADD45B ) was measured via two-step qRT-PCR. The data represent copy numbers normalised to 100,000 copies of the reference gene GAPDH (mean ± SD; n = 3 (HepG2), n = 4 (PHH Ctrl), n = 2 (PHH WD)). d The GSH/GSSG ratios of HepG2 and HepG2 ATP7B-KO cells were measured via the GSH/GSSG-Glo™ assay (means ± SDs; n = 3). e H 2 O 2 levels in HepG2 and HepG2 ATP7B-KO cells were measured by luminescence via the ROS-Glo™ H 2 O 2 assay (mean ± SD; n = 3). The effect of CuCl 2 concentrations in HepG2 or HepG2 ATP7B-KO cells (* p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001) and differences between HepG2 and HepG2 ATP7B-KO cells after 6–24 h (# p < 0.05; ## p < 0.01; ### p < 0.001; ####; p < 0.0001) were measured by two two-way ANOVA tests and a Tukey’s multiple comparison tests. Differences in gene expression between PHH Ctrl and WD cells was tested by an unpaired t-test (× p < 0.05; ×× p < 0.01). Ctrl control, PHH primary human hepatocytes, WD Wilson’s disease

Article Snippet: Cell viability of CuCl 2 (Merck) or CuSO 4 (Merck) -treated HepG2 and HepG2 ATP7B-KO cells was measurement by cell counting assay (CCK-8, Sigma-Aldrich).

Techniques: Gene Expression, Quantitative RT-PCR, Glo Assay, Comparison, Control

Journal: Molecular and Cellular Biochemistry

Article Title: High copper levels induce oxidative stress and inflammatory processes in a cell culture model of Wilson’s disease

doi: 10.1007/s11010-026-05481-6

Figure Lengend Snippet: Copper induces NFKB- and AP1-related inflammatory responses. HepG2 and HepG2 ATP7B-KO cells were treated with 0.2-0.2.6mmol/l CuCl 2 for 6 and 24 h. RNA from HepG2 and HepG2 ATP7B-KO cells and untreated PHH (Ctrl and WD) was extracted, and the gene expression of genes related to NFKB and AP1 ( a , FOS ; b , PLA2G2A ; c , JUN; d , JUNB ) was measured via two-step qRT‒PCR (mean ± SD; n = 3 (HepG2), n = 4 (PHH Ctrl), n = 2 (PHH WD)). Supernatants from HepG2 and HepG2 ATP7B-KO cells treated with CuCl 2 were collected, and various cytokine levels ( e , IL1B; f , TNF; g , IL8; h , GM-CSF) were measured via the LEGENDplex™ Assay (mean ± SD, n = 3). HepG2 and HepG2 ATP7B-KO cells were transfected with luciferase-based reporter plasmids pAP1-Luc ( i , j ), pNFKB-Luc ( k , l ) and pTA-Luc as a control and incubated with CuCl 2 . Measurement of NFKB and AP1 promoter activity was performed via a luciferase reporter assay (means ± SDs, n = 3). The effect of CuCl 2 concentrations in HepG2 or HepG2 ATP7B-KO cells (* p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001) and differences between HepG2 and HepG2 ATP7B-KO cells after 6–24 h (# p < 0.05; ## p < 0.01; ### p < 0.001; #### p < 0.0001) were measured by two-way ANOVA tests and Tukey’s multiple comparison tests. Differences in gene expression between PHH Ctrl and WD cells were tested by an unpaired t-test (× p < 0.05; ×× p < 0.01; ×××× p < 0.0001). Ctrl control, ns not significant, PHH primary human hepatocytes, WD Wilson’s disease

Article Snippet: Cell viability of CuCl 2 (Merck) or CuSO 4 (Merck) -treated HepG2 and HepG2 ATP7B-KO cells was measurement by cell counting assay (CCK-8, Sigma-Aldrich).

Techniques: Gene Expression, Transfection, Luciferase, Control, Incubation, Activity Assay, Reporter Assay, Comparison