human hepatocytes  (ATCC)

Journal: One Health

Article Title: Human 3D liver spheroids support productive infection of a novel tick-borne phenuivirus

doi: 10.1016/j.onehlt.2026.101321

Figure Lengend Snippet: Assembly and characterization of human 3D liver spheroids via DNA origami NAC-linkers. (A) Schematic of 3D liver spheroid self-assembly from primary human hepatocytes, liver sinusoidal endothelial cells, and Kupffer cells using NAC-linkers. (B) Atomic force microscopy image of NAC-linkers. Scale bars, 200 nm. (C) 1% agarose gel electrophoresis confirming cholesterol-modified NAC-linkers assembly (lanes: DNA marker, M13mp18 scaffold, and NAC-linkers). (D) Bright-field image of a mature spheroid. (E) Hematoxylin and eosin (H&E) staining of a spheroid section. (F) Immunofluorescence staining of cell type markers in human 3D liver spheroids: albumin (ALB, hepatocytes), CD31 (endothelial cells), and CD68 (Kupffer cells). Scale bars, 200 μm.

Article Snippet: Primary human hepatocytes, liver sinusoidal endothelial cells, and Kupffer cells (IxCell Biotechnology) were mixed at specific ratios and co-incubated with NAC-Linker A and B (Puheng Biomedicine, NAC001) to facilitate NAC structure formation on the cell surfaces.

Techniques: Microscopy, Agarose Gel Electrophoresis, Modification, Marker, Staining, Immunofluorescence

Journal: One Health

Article Title: Human 3D liver spheroids support productive infection of a novel tick-borne phenuivirus

doi: 10.1016/j.onehlt.2026.101321

Figure Lengend Snippet: Adaptation and pathogenesis of MKWV in human 3D liver spheroids. (A) Schematic of serial passaging of the HLJ1 strain in spheroids, yielding the adapted NAC-Org5 strain. (B, C) Viral RNA copies (B) and TCID₅₀ titers (C) across passages (P1-P5). (D) Bright-field image of spheroids infected with passage 5 (P5) virus, showing structural disruption. Scale bar, 100 μm. (E) Quantification of spheroid diameter post-infection. (F) Transmission electron micrographs of virions within cytoplasmic vesicles of infected spheroids. Scale bars: 1 μm (left), 200 nm (right). (G) Representative images and quantification of nuclei showing infection-induced cell death. Scale bar, 200 μm. (H) Western blot detecting cleaved caspase-3 in spheroids at 48 and 72 h post-infection (hpi). (I) Multiplex immunofluorescence showing NAC-Org5 tropism for CD31 + endothelial cells and CD68 + Kupffer cells, with weaker detection in ALB + hepatocytes. Scale bar, 200 μm. (J) Functional assessment of infected spheroids: ATP (viability), ALT/AST/LDH (damage), ALB/urea (synthetic function). (K) RT-qPCR analysis of pro-inflammatory cytokine mRNA expression, normalized to β-actin. Data are mean ± SD ( n = 5 biological replicates). * p < 0.05, ** p < 0.01.

Article Snippet: Primary human hepatocytes, liver sinusoidal endothelial cells, and Kupffer cells (IxCell Biotechnology) were mixed at specific ratios and co-incubated with NAC-Linker A and B (Puheng Biomedicine, NAC001) to facilitate NAC structure formation on the cell surfaces.

Techniques: Passaging, Infection, Virus, Disruption, Transmission Assay, Western Blot, Multiplex Assay, Immunofluorescence, Functional Assay, Quantitative RT-PCR, Expressing

Journal: One Health

Article Title: Human 3D liver spheroids support productive infection of a novel tick-borne phenuivirus

doi: 10.1016/j.onehlt.2026.101321

Figure Lengend Snippet: Pathogenicity of the NAC-Org5 strain in murine models. (A) Experimental schematic for intracranial (3-day-old) and intraperitoneal (3-week-old) inoculation of BALB/c mice. (B, C) Survival (B) and weight change (C) of suckling mice after NAC-Org5 infection. (D) Viral load in tissues and blood of suckling mice at 7 dpi. (E, F) Survival (E) and weight change (F) of 3-week-old mice. (G) Viral load in tissues and blood of 3-week-old mice at 7 dpi. Data are from 3 independent experiments. (H) Representative H& E -stained liver sections from 3-week-old mice at 7 and 15 dpi, showing inflammatory infiltrates and hepatocyte necrosis that resolves by 15 dpi. Scale bar, 100 μm. *** p < 0.001.

Article Snippet: Primary human hepatocytes, liver sinusoidal endothelial cells, and Kupffer cells (IxCell Biotechnology) were mixed at specific ratios and co-incubated with NAC-Linker A and B (Puheng Biomedicine, NAC001) to facilitate NAC structure formation on the cell surfaces.

Techniques: Infection, Staining

Journal: Frontiers in Pharmacology

Article Title: Downregulation of microRNA-300-3p promotes steatosis-to-MASH progression by regulating STX17

doi: 10.3389/fphar.2026.1804415

Figure Lengend Snippet: MiR-300-3p was decreased in the livers of MAFLD mice and FFA-induced HepG2 cells. (A) MiR-300-3p was decreased in the livers of MAFLD mice. (B) MiR-300-3p was downregulated in the FFA-induced HepG2 cells. (C) MiR-300-3p was downregulated in HepG2 cells successfully. (D) MiR-300-3p was overexpressed in HepG2 cells successfully. Data in (A–D) are means±SDs (n = 3). *P < 0.05, **P < 0.01. NC, Healthy control; OC, over-expression control; IC, inhibition control.

Article Snippet: The human hepatocyte cell line HepG2 was procured from the American Type Culture Collection (ATCC, United States) and cultivated in a humidified incubator maintained at 37 °C with a 5% CO 2 atmosphere.

Techniques: Control, Over Expression, Inhibition

Journal: Frontiers in Pharmacology

Article Title: Downregulation of microRNA-300-3p promotes steatosis-to-MASH progression by regulating STX17

doi: 10.3389/fphar.2026.1804415

Figure Lengend Snippet: Downregulation of miR-300-3p promotes FFA-induced lipid accumulation and hepatic inflammation in HepG2 cells. (A) The experimental flow of the MAFLD cell model construction and miR-300-3p mimics, miR-300-3p inhibitors and as well as corresponding scrambled controls transfection in HepG2 cells. (B,C) Intercellular TG contents in miR-300-3p -inhibited and -overexpressing in HepG2 cells. (D) Oil Red O staining (400×) and the relative areas of lipid droplets in miR-300-3p-inhibited and -overexpressing in HepG2 cells. (E,F) mRNA levels of FASN, SREBP-1c, IL-6, IL-2 and TNF-αin miR-300-3p -inhibited and -overexpressing in HepG2 cells. (G) Protein levels of FASN, SREBP-1c and TNF-α in miR-300-3p -inhibited and -overexpressing in HepG2 cells. Data in (B,C) and (E–G) are means±SDs (n = 3). *P < 0.05, **P < 0.01, *** P < 0.001, **** P < 0.0001. OC, over-expression control; IC, inhibition control.

Article Snippet: The human hepatocyte cell line HepG2 was procured from the American Type Culture Collection (ATCC, United States) and cultivated in a humidified incubator maintained at 37 °C with a 5% CO 2 atmosphere.

Techniques: Transfection, Staining, Over Expression, Control, Inhibition

Journal: Frontiers in Pharmacology

Article Title: Downregulation of microRNA-300-3p promotes steatosis-to-MASH progression by regulating STX17

doi: 10.3389/fphar.2026.1804415

Figure Lengend Snippet: Downregulation of miR-300-3p induces apoptosis in HepG2 cells to promote it. (A,B) The apoptosis rate in miR-300-3p -inhibited and -overexpressing in HepG2 cells. (C) mRNA levels of Bax, Bcl-2 and Caspase-3 in miR-300-3p -inhibited and -overexpressing in HepG2 cells. (D) Protein levels of Bax, Bcl-2 and Caspase-3 in miR-300-3p -overexpressing and -inhibited in HepG2 cells. Data in (A–D) are means±SDs (n = 3). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. OC, over-expression control; IC, inhibition control.

Article Snippet: The human hepatocyte cell line HepG2 was procured from the American Type Culture Collection (ATCC, United States) and cultivated in a humidified incubator maintained at 37 °C with a 5% CO 2 atmosphere.

Techniques: Over Expression, Control, Inhibition

Journal: Frontiers in Pharmacology

Article Title: Downregulation of microRNA-300-3p promotes steatosis-to-MASH progression by regulating STX17

doi: 10.3389/fphar.2026.1804415

Figure Lengend Snippet: MiR-300-3p regulates STX17 in HepG2 cells. (A,B) GO enrichment analysis (Biological Process category) and KEGG enrichment analysis diagrams of sixteen predicted target genes of miR-300-3p. (C) The predicted binding sites between miR-300-3p and STX17 were putatively identified via the TargetScan database. (D) Dual-luciferase reporter assay (DLRA) in HepG2 cells validated that STX17 is a direct target gene of miR-300-3p. (E,F) The mRNA and protein expression levels of STX17 in HepG2 cells with miR-300-3p inhibition or overexpression. Data in (A–D) are means±SDs (n = 3). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. OC, overexpression control; IC, inhibition control.

Article Snippet: The human hepatocyte cell line HepG2 was procured from the American Type Culture Collection (ATCC, United States) and cultivated in a humidified incubator maintained at 37 °C with a 5% CO 2 atmosphere.

Techniques: Binding Assay, Luciferase, Reporter Assay, Expressing, Inhibition, Over Expression, Control

Journal: Frontiers in Pharmacology

Article Title: Downregulation of microRNA-300-3p promotes steatosis-to-MASH progression by regulating STX17

doi: 10.3389/fphar.2026.1804415

Figure Lengend Snippet: P62 and LC3-II in miR-300-3p -inhibited and -overexpressing in HepG2 cells. (A) The protein levels of P62 and LC3-II in miR-300-3p -inhibited and -overexpressing in HepG2 cells. (B) P62 and LC3-II in miR-300-3p -inhibited and -overexpressing by Immunofluorescence assay. Data in (A-D) are means±SDs (n = 3). *P < 0.05, **P < 0.01,***P < 0.001,****P < 0.0001. OC, overexpression control; IC, inhibition control.

Article Snippet: The human hepatocyte cell line HepG2 was procured from the American Type Culture Collection (ATCC, United States) and cultivated in a humidified incubator maintained at 37 °C with a 5% CO 2 atmosphere.

Techniques: Immunofluorescence, Over Expression, Control, Inhibition

Journal: Frontiers in Pharmacology

Article Title: Downregulation of microRNA-300-3p promotes steatosis-to-MASH progression by regulating STX17

doi: 10.3389/fphar.2026.1804415

Figure Lengend Snippet: MiR-300-3p promoted autophagy by regulating STX17, reduces lipid accumulation, inflammatory response, and decreases hepatocyte apoptosis. (A) STX17 was inhibited or overexpressed in HepG2 cells successfully. (B) Protein levels of FASN, SREBP-1c and TNF-αin STX17 -inhibited and -overexpressing in HepG2 cells. (C,D) Intercellular TG contents in STX17-inhibited and -overexpressing in HepG2 cells. (E) Oil Red O staining (400×) and relative areas of lipid droplets in STX17-inhibited and -STX17 in HepG2 cells. (F) Protein levels of Bax, Capase-3 and Bcl-2 in STX17 -inhibited and -overexpressing in HepG2 cells. Data in (A–D) and (F) are means±SDs (n = 3). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. OC, miR-300-3p overexpression control; IC,miR-300-3p inhibition control; Control, STX17-overexpression control or STX17-inhibition control; Si-STX17, STX17-inhibition; Over-STX17, STX17-overexpression.

Article Snippet: The human hepatocyte cell line HepG2 was procured from the American Type Culture Collection (ATCC, United States) and cultivated in a humidified incubator maintained at 37 °C with a 5% CO 2 atmosphere.

Techniques: Staining, Over Expression, Control, Inhibition

Journal: Frontiers in Pharmacology

Article Title: Downregulation of microRNA-300-3p promotes steatosis-to-MASH progression by regulating STX17

doi: 10.3389/fphar.2026.1804415

Figure Lengend Snippet: P62, LC3-II in STX17-inhibited and -overexpressing in HepG2 cells. (A) Protein levels of P62, LC3-II in STX17-inhibited and -overexpressing in HepG2 cells. (B) P62 and LC3-II in STX17-inhibited and -overexpressing by Immunofluorescence assay. Data in (A-D) are means±SDs (n = 3). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. OC, overexpression control; IC, inhibition control.

Article Snippet: The human hepatocyte cell line HepG2 was procured from the American Type Culture Collection (ATCC, United States) and cultivated in a humidified incubator maintained at 37 °C with a 5% CO 2 atmosphere.

Techniques: Immunofluorescence, Over Expression, Control, Inhibition

Journal: Immunity, Inflammation and Disease

Article Title: Asperosaponin VI Alleviates Cisplatin‐Induced Liver Injury Through the Nrf2/HO‐1 Signaling Pathway

doi: 10.1002/iid3.70454

Figure Lengend Snippet: Protective efficacy of AVI against cisplatin‐induced cytotoxicity and apoptosis in LO2 cells. (A) Indicates that LO2 cell viability was not affected by AVI intervention at concentrations ranging from 0 to 400 μM over a period of 2 days. (B) Indicates a significant difference in LO2 cell viability following cisplatin treatment with vs. without AVI co‐culture. (C) Illustrates the dose‐response curves of cell viability to cisplatin following different pretreatment durations (0, 6, 12, and 24 h) with AVI. (D) Shows the apoptosis rate in LO2 cells was quantified using Annexin V‐FITC double staining, (E) shows Cis group has the highest rate of cell apoptosis, while co‐culture with AVI significantly reduced the overall cell apoptosis rate caused by Cis. (F) Shows the apoptosis of cells in each group. Apoptosis was significantly observed in the Cis group, while AVI co‐culture could significantly improve apoptosis. (G) Shows the total amount of ROS produced by cells in each group. The Cis group significantly increased ROS, while AVI co‐culture could significantly reduce the generation of ROS within cells. ( n = 3, * p < 0.05, ** p < 0.01,*** p < 0.001).

Article Snippet: Human LO2 hepatocytes (Procell Biotechnology, China; catalog CM‐0111) were cultured in complete growth medium supplemented with 10% fetal bovine serum (FBS; Biosharp, China, catalog BL201A) and 1% penicillin‐streptomycin (Biosharp, China, catalogBL505A).

Techniques: Co-Culture Assay, Double Staining, Produced

Journal: Immunity, Inflammation and Disease

Article Title: Asperosaponin VI Alleviates Cisplatin‐Induced Liver Injury Through the Nrf2/HO‐1 Signaling Pathway

doi: 10.1002/iid3.70454

Figure Lengend Snippet: Transcriptome sequencing results from four experimental groups of LO2 cells. (A) Principal component analysis (PCA) shows clear separation along PC1 among different treatment groups, indicating value for further analysis. (B) Gene expression distribution (violin plot) demonstrates consistency in expression levels across samples. (C) Heatmap of differentially expressed gene clustering, and (D) Volcano plot of differentially expressed genes, both reveal significant differential gene expression between the Cis group and the AVI+Cis group. (E) Gene Ontology (GO) enrichment analysis shows that the gene set is significantly enriched in the three categories: biological process, molecular function, and cellular component. (F) Based on KEGG pathway enrichment analysis results, the gene set primarily covers key physiological and pathological processes such as immune defense, apoptosis, and signal transduction. (G) Results of Gene Set Enrichment Analysis (GSEA) show that gene sets involved in oxidative stress, inflammatory response, and programmed cell death are significantly negatively enriched in AVI+Cis group.

Article Snippet: Human LO2 hepatocytes (Procell Biotechnology, China; catalog CM‐0111) were cultured in complete growth medium supplemented with 10% fetal bovine serum (FBS; Biosharp, China, catalog BL201A) and 1% penicillin‐streptomycin (Biosharp, China, catalogBL505A).

Techniques: Sequencing, Gene Expression, Expressing, Transduction

Journal: Immunity, Inflammation and Disease

Article Title: Asperosaponin VI Alleviates Cisplatin‐Induced Liver Injury Through the Nrf2/HO‐1 Signaling Pathway

doi: 10.1002/iid3.70454

Figure Lengend Snippet: AVI suppresses hepatocyte inflammation and apoptosis in LO2 cells via activation of the Nrf2/HO‐1 axis. Total protein was extracted from the LO2cell from the control, Cis, and AVI+Cis, Bru+AVI+Cis, Vitc+Cis groups. (A–C) Representative images of western blots depicting the levels of Nrf2 and HO‐1 in the LO2 cell, and the protein/GAPDH ratios determined by densitometric analysis of the western blots. (D–H) Representative images of western blots depicting the levels of Caspase‐1, NF‐κB, NLRP3 and Caspase‐3 in the LO2 cell, and the protein/GAPDH ratios determined by densitometric analysis of the western blots.

Article Snippet: Human LO2 hepatocytes (Procell Biotechnology, China; catalog CM‐0111) were cultured in complete growth medium supplemented with 10% fetal bovine serum (FBS; Biosharp, China, catalog BL201A) and 1% penicillin‐streptomycin (Biosharp, China, catalogBL505A).

Techniques: Activation Assay, Control, Western Blot

Journal: Molecules

Article Title: Lutein Modulates Stress-Responsive Signaling Pathways in THLE-2 Human Hepatocytes Under Intestinal Failure–Associated Liver Disease Conditions

doi: 10.3390/molecules31091413

Figure Lengend Snippet: Effect of lipopolysaccharide (LPS) and Intralipid on the metabolic activity of THLE-2 cells after 24 h of incubation. Cells were treated with a range of concentrations of LPS ( A ) or Intralipid ( B ), and metabolic activity was assessed using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Results are expressed as a percentage relative to untreated control cells (set as 100%). Data are presented as mean ± SEM ( n = 3).

Article Snippet: Human immortalized hepatocytes THLE-2 (ATCC CRL-2706) were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Activity Assay, Incubation, MTT Assay, Control

Journal: Molecules

Article Title: Lutein Modulates Stress-Responsive Signaling Pathways in THLE-2 Human Hepatocytes Under Intestinal Failure–Associated Liver Disease Conditions

doi: 10.3390/molecules31091413

Figure Lengend Snippet: Effects of LPS (0.1 µg/mL), Intralipid (INT, 10 mg/mL), or their combination (IFALD model) on signaling pathways in THLE-2 cells after 24 h of exposure. Total and phosphorylated levels of p38, extracellular signal–regulated kinases 1/2 (ERK1/2), c-Jun N-terminal kinase (JNK), nuclear factor κB (NF-κB), and protein kinase B (Akt) were quantified based on fluorescence intensity using the MAGPIX system. Data are expressed as fold change relative to untreated controls (mean ± SEM, n = 3). Statistical significance was determined using Dunnett’s test. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001 vs. control. The horizontal dotted line indicates the control levels.

Article Snippet: Human immortalized hepatocytes THLE-2 (ATCC CRL-2706) were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Protein-Protein interactions, Fluorescence, Control

Journal: Molecules

Article Title: Lutein Modulates Stress-Responsive Signaling Pathways in THLE-2 Human Hepatocytes Under Intestinal Failure–Associated Liver Disease Conditions

doi: 10.3390/molecules31091413

Figure Lengend Snippet: Effects of LPS (0.1 µg/mL), Intralipid (INT, 10 mg/mL), or their combination (IFALD model) on signaling pathways in THLE-2 cells after 24 h of exposure. Total and phosphorylated levels of signal transducer and activator of transcription 3 and 5 (STAT3 and STAT5), cAMP response element-binding protein (CREB), and 70 kDa ribosomal protein S6 kinase (p70S6K) were quantified based on fluorescence intensity using the MAGPIX system. Data are expressed as fold change relative to untreated controls (mean ± SEM, n = 3). Statistical significance was determined using Dunnett’s test. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001 vs. control. The horizontal dotted line indicates the control levels.

Article Snippet: Human immortalized hepatocytes THLE-2 (ATCC CRL-2706) were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Protein-Protein interactions, Binding Assay, Fluorescence, Control

Journal: Molecules

Article Title: Lutein Modulates Stress-Responsive Signaling Pathways in THLE-2 Human Hepatocytes Under Intestinal Failure–Associated Liver Disease Conditions

doi: 10.3390/molecules31091413

Figure Lengend Snippet: Effect of lutein on THLE-2 cell metabolic activity after 24 h of exposure. Cell metabolic activity was assessed using the MTT assay across a range of lutein concentrations (1–100 µM) and expressed as a percentage relative to untreated control cells. Data are presented as mean ± SEM ( n = 3).

Article Snippet: Human immortalized hepatocytes THLE-2 (ATCC CRL-2706) were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Activity Assay, MTT Assay, Control

Journal: Molecules

Article Title: Lutein Modulates Stress-Responsive Signaling Pathways in THLE-2 Human Hepatocytes Under Intestinal Failure–Associated Liver Disease Conditions

doi: 10.3390/molecules31091413

Figure Lengend Snippet: Effects of lutein on cellular signaling components in the in vitro IFALD model. THLE-2 cells were treated with LPS (0.1 µg/mL) and Intralipid (10 mg/mL) to mimic IFALD-like conditions and co-treated with lutein (LUT) at concentrations of 10 or 25 µM for 24 h. Total and phosphorylated levels of p38, ERK1/2, JNK, NF-κB, Akt, and STAT5 were quantified based on mean fluorescence intensity using the MAGPIX System. Data are expressed as fold change relative to untreated control cells (mean ± SEM, n = 3). Statistical significance was determined using Dunnett’s test. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001 vs. control; # p < 0.05, ## p < 0.01, and ### p < 0.001 vs. IFALD.

Article Snippet: Human immortalized hepatocytes THLE-2 (ATCC CRL-2706) were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: In Vitro, Fluorescence, Control

Journal: Molecules

Article Title: Lutein Modulates Stress-Responsive Signaling Pathways in THLE-2 Human Hepatocytes Under Intestinal Failure–Associated Liver Disease Conditions

doi: 10.3390/molecules31091413

Figure Lengend Snippet: Effects of lutein on cellular signaling components in the in vitro IFALD model. THLE-2 cells were treated with LPS (0.1 µg/mL) and Intralipid (10 mg/mL) to mimic IFALD-like conditions and co-treated with lutein (LUT) at concentrations of 10 or 25 µM for 24 h. ( A ) Total and phosphorylated levels of STAT3, CREB, and p70S6K, quantified based on mean fluorescence intensity using the MAGPIX System. ( B ) mRNA expression levels of sterol regulatory element-binding protein 2 ( SREBF2 ), ATP-binding cassette subfamily A member 1 ( ABCA1 ), AMP-activated protein kinase α2 ( PRKAA2 ), cholesterol 7 α-hydroxylase ( CYP7A1 ), and 3-hydroxy-3-methylglutaryl-CoA reductase ( HMGCR ), determined by quantitative real-time polymerase chain reaction (qPCR). Data are expressed as fold change relative to untreated control cells (mean ± SEM, n = 3). Statistical significance was determined using Dunnett’s test. * p < 0.05, ** p < 0.01, **** p < 0.0001 vs. control; # p < 0.05, ## p < 0.01, and #### p < 0.0001 vs. IFALD.

Article Snippet: Human immortalized hepatocytes THLE-2 (ATCC CRL-2706) were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: In Vitro, Fluorescence, Expressing, Binding Assay, Real-time Polymerase Chain Reaction, Control

Journal: Molecules

Article Title: Lutein Modulates Stress-Responsive Signaling Pathways in THLE-2 Human Hepatocytes Under Intestinal Failure–Associated Liver Disease Conditions

doi: 10.3390/molecules31091413

Figure Lengend Snippet: Proposed mechanism of action of lutein in THLE-2 hepatocytes exposed to IFALD-related triggers—LPS and omega-6-rich Intralipid. AA, arachidonic acid; TLR4, Toll-like receptor 4; IRS1, insulin receptor substrate 1; PI3K, phosphoinositide 3-kinase; IL-1β, interleukin-1β; COX-2, cyclooxygenase-2; IL-6, interleukin-6; HDL, high-density lipoprotein; FAs, fatty acids.

Article Snippet: Human immortalized hepatocytes THLE-2 (ATCC CRL-2706) were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques:

Journal: Molecules

Article Title: Lutein Modulates Stress-Responsive Signaling Pathways in THLE-2 Human Hepatocytes Under Intestinal Failure–Associated Liver Disease Conditions

doi: 10.3390/molecules31091413

Figure Lengend Snippet: Effect of free lutein, AlbLuteN, and blank albumin nanosuspension on the metabolic activity of THLE-2 human hepatocytes after 48 h of exposure, assessed by the MTT assay. Free lutein and AlbLuteN were tested at equivalent lutein concentrations (1–100 µM), while the blank nanosuspension was applied at carrier concentrations matching those in AlbLuteN. Cell metabolic activity is expressed as a percentage of the untreated control. Data are presented as mean ± SEM ( n = 3).

Article Snippet: Human immortalized hepatocytes THLE-2 (ATCC CRL-2706) were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Activity Assay, MTT Assay, Control

Journal: bioRxiv

Article Title: Inhibition of p107 alleviates liver steatosis by reducing de novo fatty acid synthesis

doi: 10.64898/2026.04.14.718271

Figure Lengend Snippet: (A) p107 protein levels in THLE2 cells transfected with siRNA p107 or siRNA control for 48 hours (n=3 per group). (B) Representative microphotographs of Oil Red O staining (left pannel) of THLE2 cells downregulating p107 (sip107) for 48 hours (n = 3 per group) and Oil Red O semiquantification (right pannel) (n = 3 per group). (C) Quantification of immunoblot analysis of de novo lipogenesis markers (n = 3 per group) and a representative immunoblot. (D) RNA expression of de novo lipogenesis markers (n = 3 per group. (E) De novo synthesis of free fatty acids (FFA) and triglycerides (TG) in THLE2 (n = 6 per group). (F) FAO activity in THLE2 cells transfected with siRNA p107 or siRNA control for 48 hours (n=5 per group). (G) Oxidation rate of palmitic acid (n = 6 per group). (H and I)) OCR and (I) ECAR of p107-silenced THLE2 (n = 17-19 per group). (J) Basal energetic metabolic states, based on quantification of ECAR and OCR during basal metabolism. (K) p107 protein levels in THLE2 after overexpressing p107 (n = 3 per group). (L) Representative microphotographs (left pannel) and semiquantification (right pannel) of Oil Red O staining of THLE2 cells overexpressing p107 (plasmid p107) for 24 hours. Oil Red O staining was quantified using ImageJ and normalized to the total number of nuclei per field (n = 4 per group). (M) Quantification of immunoblot analysis of de novo lipogenesis markers (n = 3 per group) and a representative immunoblot. (N) OCR of p107-overexpressed THLE2 (n =10 per group). GAPDH was used to normalize protein levels. Data are expressed as mean ±SEM. *p < 0.05, **p < 0.01, ***p <0.001, using a Student’s t test .

Article Snippet: THLE2 human hepatic cell line (American Type Culture Collection, ATCC) was cultured in bronchial epithelial cell basal medium (BEBM) supplemented with a growth factors BulleKit (Lonza/Clonetics Corporation), 70ng/mL phosphoethanolamine, 5 ng/mL epidermal growth factor, 10% (v/v) FBS and 1% (v/v) Glutamine-Penicillin-Streptomycin solution (MERCK).

Techniques: Transfection, Control, Staining, Western Blot, RNA Expression, Activity Assay, Plasmid Preparation

Journal: bioRxiv

Article Title: Inhibition of p107 alleviates liver steatosis by reducing de novo fatty acid synthesis

doi: 10.64898/2026.04.14.718271

Figure Lengend Snippet: (A) Volcano plot of total protein expression in p107 liver KO mice compared to shLucif controls (n = 7). Red and blue points indicate significantly up- and downregulated proteins (p < 0.05). (B) Volcano plot of phosphosite abundance in p107 liver KO mice compared to shLucif controls (n = 7). Points represent individual phosphosites—annotated by their parent protein name—with red and blue indicating significant changes (p < 0.05). (C) Heatmap of total protein expression differences grouped by GO Biological Process terms, filtered by significance (p < 0.001). (D) Heatmap of robust total protein expression differences grouped by GO terms, utilizing strict filtering (> 3 combined razor and unique peptides, p < 0.001, absolute t-test difference > 0.58). (E) Quantification of p107 (left panel) and FAS (right panel) in THLE2 after overexpressing p107 (plasmid p107) and silencing FAS (siRNA FAS) during 24h (n = 3 per group) and representative immunoblot. (F) Representative microphotographs of Oil Red O staining of THLE2 cells overexpressing p107 (plasmid p107) and silencing FAS (siRNA FAS) for 24 hours. (G) Semiquantification of Oil Red O staining. Vinculin was used to normalize protein levels. Data are expressed as mean ±SEM. *p < 0.05, **p < 0.01, ***p <0.001, using a Student’s t test .

Article Snippet: THLE2 human hepatic cell line (American Type Culture Collection, ATCC) was cultured in bronchial epithelial cell basal medium (BEBM) supplemented with a growth factors BulleKit (Lonza/Clonetics Corporation), 70ng/mL phosphoethanolamine, 5 ng/mL epidermal growth factor, 10% (v/v) FBS and 1% (v/v) Glutamine-Penicillin-Streptomycin solution (MERCK).

Techniques: Expressing, Phospho-proteomics, Plasmid Preparation, Western Blot, Staining