human normal hepatocytes  (ATCC)

Journal: Journal of Cellular and Molecular Medicine

Article Title: Regulation of Autophagy and Metabolism in Hepatocellular Carcinoma: Involvement of Wnt‐β‐Catenin Pathway

doi: 10.1111/jcmm.71070

Figure Lengend Snippet: Inhibition of cell viability and colony formation, and induction of apoptosis by riluzole. (A) Expression of stem cell markers in HCC CSCs. CSCs were stained with CD44 and CD133 antibody and the expression of CD44 and CD133 was measured by immunocytochemistry. (B, C) HCC cell lines (HepG2 and Hep3B), cancer stem cells (CSCs), and human normal hepatocytes were seeded in 96‐well plates, treated with or without riluzole (0–40 μM) for 72 h, and cell viability was measured by CellTiter‐Glo Luminescent Cell Viability Assay (Promega). (D) Colony formation. HCC cell lines (HepG2, Hep3B, SNU‐382 and SNU‐475) and CSCs were treated with riluzole (0–20 μM). Number of colonies formed at 21 days was counted under a microscope. (E) Normal human hepatocytes, HCC cell lines (HepG2 and Hep3B) and CSCs were treated with riluzole (0–20 μM) for 72 h. Apoptosis was measured by TUNEL assay. Data represent mean ± SD ( n = 4). *, # and % = significantly different from control and each other; p < 0.05.

Article Snippet: Human normal hepatocytes were purchased from ATCC.

Techniques: Inhibition, Expressing, Staining, Immunocytochemistry, Cell Viability Assay, Microscopy, TUNEL Assay, Control

Journal: Biotechnology Reports

Article Title: Insulin Resistance, Anti-inflammatory, and Antioxidant In vitro Activity of Heterologously Expressed Arenin from Dryophytes arenicolor

doi: 10.1016/j.btre.2026.e00947

Figure Lengend Snippet: In vitro modulation of hepatic insulin resistance by arenin in hepatic insulin-resistant (HepG2) cells . Arenin concentrations (3.90–1000 µg/mL) were incubated for 5 h in hepatic insulin-resistant cells. Negative control (N) cells were cultured in the absence of insulin, whereas insulin-resistant (IR) control cells were cultured in the presence of insulin. Rosiglitazone (ROSI) at 10 μM (3.57 µg/mL) was used as the positive reference control. Different letters abc indicate statistical differences between concentrations analyzed using Tukey test ( p < 0.05).

Article Snippet: Human hepatocyte carcinoma (HepG2) cells, human primary dermal fibroblasts (HDFa) cells, and murine macrophage (RAW 264.7) cells were obtained from the American Type Culture Collection (ATCC, VA, USA).

Techniques: In Vitro, Incubation, Negative Control, Cell Culture, Control

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: bioRxiv

Article Title: On the role of cell chaining in the attenuation of a Listeria monocytogenes divIVA mutant

doi: 10.64898/2026.01.05.697648

Figure Lengend Snippet: (A) Contribution of divIVA , mreB and ezrA to hepatocyte invasion. Invasion of L. monocytogenes strains EGD-e (wt), BUG2214 (Δ prfA ), LMS2 (Δ divIVA ), LMSW49 (i mreB ) and LMJR183 (i ezrA ) pre-grown in the absence or presence of 1 mM IPTG into HepG2 hepatocytes. Values are expressed relative to wild type. Average values and standard deviations are shown. Asterisks mark statistically significant differences compared to wild type ( P <0.01, t -test with Bonferroni Holm correction). (B) Contribution of divIVA , mreB and ezrA to intracellular growth in macrophages. J774 mouse macrophages were infected with L. monocytogenes strains EGD-e (wt), LMS250 (Δ hly ), LMS2 (Δ divIVA ), LMSW49 (i mreB ) and LMJR183 (i ezrA ) and then cultivated in the absence or presence of IPTG, where appropriate. Average values and standard deviations were calculated from experiments performed in triplicates. Statistically significant differences (compared to wild type or between depleted and induced conditions) are marked by asterisks ( P <0.01, t -test with Bonferroni-Holm correction, n. s. – not significant). (C) Effect of DivIVA, MreB or EzrA depletion on cell-to-cell spread. Plaque formation assay using 3T3 mouse fibroblasts with L. monocyctogenes strains EGD-e (wt), LMS30 (i divIVA ), LMSW49 (i mreB ) and LMJR183 (i ezrA ) ± IPTG. (D) Necessity of DivIVA, MreB and EzrA for actin coating and actin tail formation. J774 mouse macrophages were infected with DsRed producing L. monocytogenes strains LMJD20 (wt), LMSF2 (Δ actA ), LMSW205 (Δ divIVA ), LMSW202 (EzrA depletion) and LMSW203 (MreB depletion). Infections were carried out in the absence of IPTG and analyzed by fluorescence microscopy 4 hours post infection after DAPI and phalloidin staining. Composite images are shown.

Article Snippet: Invasion of L. monocytogenes into HepG2 human hepatocytes (ATCC® HB-8065 TM ) was quantified as described earlier [ ].

Techniques: Infection, Plaque Formation Assay, Fluorescence, Microscopy, Staining

Journal: bioRxiv

Article Title: On the role of cell chaining in the attenuation of a Listeria monocytogenes divIVA mutant

doi: 10.64898/2026.01.05.697648

Figure Lengend Snippet: (A) The prfA* mutation suppresses the invasion defect of the Δ divIVA mutant. Invasion of L. monocytogenes strains EGD-e (wt), BUG2214 (Δ prfA ), BUG3057 ( prfA* ), LMS2 (Δ divIVA ) and LMSW251 (Δ divIVA prfA* ) into HepG2 hepatocytes. Values are expressed relative to wild type. Average values and standard deviations are shown. The asterisks mark statistically significant differences compared to wild type ( P <0.01, t -test with Bonferroni-Holm correction) or to the Δ divIVA prfA* mutant ( P <0.01, t -test, n. s. – not significant). (B) No suppression of the intracellular replication defect of the Δ divIVA mutant by the prfA* mutation. J774 mouse macrophages were infected with the same strains as in panel A. Average values and standard deviations were calculated from experiments performed in triplicates. No statistically significant difference in the number of intracellular bacteria was detected between Δ divIVA and Δ divIVA prfA* cells six hours post infection ( P <0.01, t -test, n. s. – not significant). (C) Dominance of the Δ divIVA deletion over the prfA* mutation in a plaque forming assay. The same strains as above were used to infect 3T3 mouse fibroblasts to analyze cell-to-cell spread.

Article Snippet: Invasion of L. monocytogenes into HepG2 human hepatocytes (ATCC® HB-8065 TM ) was quantified as described earlier [ ].

Techniques: Mutagenesis, Infection, Bacteria

Journal: bioRxiv

Article Title: On the role of cell chaining in the attenuation of a Listeria monocytogenes divIVA mutant

doi: 10.64898/2026.01.05.697648

Figure Lengend Snippet: (A) Localisation of Δ divIVA suppressor mutations M95I and T123A in the SecA2 protein. SecA2 domains (colored) and walker A and B motifs (black) are indicated. Abbreviations: NBD –nucleotide binding domain, PPXD – preprotein crosslinking domain, IRA 1/2 – intramolecular regulator of ATPase domains 1 and 2, SD – scaffold domain. (B) Micrographs showing the cellular morphology of L. monocytogenes strains EGD-e (wt), LMS2 (Δ divIVA ), LMSW222 (Δ divIVA secA2 M95I ) and LMSW223 (Δ divIVA secA2 T123A ) during growth in BHI broth at 37°C. Membranes were stained with nile red. Scale bar is 2 µm. (C) Separation of secretome samples of the same set of strains by SDS-PAGE. The position of p60 (CwhA) is indicated. (D) Flagellar motility of the same set of strains on swarming agar after two days of incubation at 30°C. (E) Cell-to-cell spread of the same strains in 3T3 mouse fibroblast. (F) Full suppression of the Δ divIVA invasion defect by secA2 mutations. The same set of strains as in panel A was used to infect HepG2 hepatocytes. Values are expressed relative to wild type. Average values and standard deviations are shown. The asterisk marks a statistically significant difference ( P <0.01, t -test with Bonferroni-Holm correction, n. s. – not significant). (G) Partial suppression of the intracellular replication defect of the Δ divIVA mutant by secA2 mutations. J774 mouse macrophages were infected with the same strains as above and intracellular bacteria were enumerated right after infection (0 h p. i.) and six hours later (6 h p. i.). Average values and standard deviations were calculated from experiments performed in triplicates. Statistically significant differences are indicated by asterisks ( P <0.01, t -test with Bonferroni-Holm correction).

Article Snippet: Invasion of L. monocytogenes into HepG2 human hepatocytes (ATCC® HB-8065 TM ) was quantified as described earlier [ ].

Techniques: Binding Assay, Staining, SDS Page, Incubation, Mutagenesis, Infection, Bacteria