mouse normal liver cell line aml12  (ATCC)

Journal: Discover Oncology

Article Title: Isosilybin B: a potential novel therapeutic agent with hepatoprotective, anticancer and antifibrotic properties

doi: 10.1007/s12672-025-03380-8

Figure Lengend Snippet: Effect of IB/SB/SM on cytotoxicity and cell cycle arrest of liver cells. A : Chemical structure of IB and SB (silybin A and silybin B in ratio 1:1). Cytotoxicity in Hepa1-6 ( B ), HepG2 ( C ) or AML12 ( D ) liver cells after 24 h treatment with IB/SB/SM. Changes in the cell cycle in Hepa1-6 ( E ), HepG2 ( F ), or AML12 ( G ) liver cells after 24 h treatment with 31.3 µg/mL of IB/SB/SM. H : Cell distribution of diploid cells in the phases of the cell cycle shown for IB (31.3 µg/mL). The data are given as means ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: Mouse normal liver hepatocyte cell line AML12 (RRID: CVCL_0140, ATCC, USA) was cultivated in DMEM/F12 medium supplemented with 10% FBS, 1% penicillin-streptomycin, 1X Insulin-Transferrin-Selenium and 40 ng/mL dexamethasone.

Techniques:

Journal: Discover Oncology

Article Title: Isosilybin B: a potential novel therapeutic agent with hepatoprotective, anticancer and antifibrotic properties

doi: 10.1007/s12672-025-03380-8

Figure Lengend Snippet: Effect of IB/SB/SM on pro-fibrotic gene expression, antioxidant activity, and ALT concentration in AML12 cells. (A-C) Expression of pro-fibrotic genes fibronectin ( Fn1 ), smooth muscle actin ( Acta2 ) and collagen I ( Col1a1 ) after 24 h treatment with TGF-β1 (10 ng/mL) and IB/SB/SM (7.8–31.3 µg/mL) in AML12 cells. (D) Western blot analysis of fibronectin expression in AML12 cells stimulated for 24 h with TGF-β1 (10 ng/mL) and IB/SB/SM at concentration 31.3 µg/mL. Representative western blot bands are shown below the graphs. (E) DPPH antioxidant activity assay. The ability of IB/SB/SM to scavenge DPPH radicals was evaluated at various concentrations (0–500 µg/mL). (F) Determination of alanine aminotransferase (ALT) concentration in the supernatants of AML 12 cells stimulated with TGF-β1 (10 ng/mL) in the presence of the tested compounds (IB, SB, SM) at different concentrations. All results are expressed as mean ± SD, * p < 0.05, ** p < 0.01, *** p < 0,001 (for western blot: vs. TGF-β1 control)

Article Snippet: Mouse normal liver hepatocyte cell line AML12 (RRID: CVCL_0140, ATCC, USA) was cultivated in DMEM/F12 medium supplemented with 10% FBS, 1% penicillin-streptomycin, 1X Insulin-Transferrin-Selenium and 40 ng/mL dexamethasone.

Techniques: Gene Expression, Antioxidant Activity Assay, Concentration Assay, Expressing, Western Blot, Control

Journal: Acta biochimica et biophysica Sinica

Article Title: Modulation of ferroptosis via YY1-SLC7A11 axis in hepatic ischemia-reperfusion injury pathogenesis.

doi: 10.3724/abbs.2025093

Figure Lengend Snippet: Figure 2. Impact of YY1 overexpression on in vitro and in vivo hepatic IRI (A) Western blot analysis of AML12 YY1 expression post IRI-OGD/R treatment. (B) Flow cytometry assessment of the effect of YY1 overexpression on IRI-OGD/R-induced apoptosis, with representative results on the left and corresponding bar graph on the right. (C) Evaluation of liver tissue damage in Sham and IRI-I/R treated mice following YY1 overexpression, with representative H&E staining results on the left showing a low magnification view, white dashed lines indicating the injury area (scale bar = 100 μm), and local enlargement of the injury area (scale bar = 25 μm), and Suzuki scores on the right. (D) ELISA results of serum ALT and AST levels in each group of mice. (E,F) Expression levels of the inflammatory factors IL-1β, TNF-α, and MCP1 in mouse serum and liver tissue within each group assessed by ELISA and RT-qPCR. (G) Expression levels of the chemokines CXCl2 and CCL5 in mouse liver tissues in each group measured by RT-qPCR. (H) Immunohistochemistry was performed to evaluate the infiltration of Ly6G+ neutrophils in the liver tissues of different groups of mice. A representative image is presented on the left, accompanied by a corresponding statistical bar graph on the right (scale bar = 50 μm). n = 10 in each group for the animal experiments. Cell experiments were repeated three times. *P < 0.05, **P < 0.01, ***P < 0.001. ns, indicates no significance.

Article Snippet: The AML12 normal mouse liver cell line (CRL2254; ATCC, Manassas, USA) was cultured in DMEM/F-12 (11320033; Thermo Fisher Scientific) supplemented with 10 μg/mL insulin, 5.5 μg/mL transferrin, 5 ng/mL selenium, 40 ng/mL dexamethasone, 10% FBS (10100147C; Thermo Fisher Scientific), and 1% penicillin-streptomycin (15140163; Thermo Fisher Scientific).

Techniques: Over Expression, In Vitro, In Vivo, Western Blot, Expressing, Flow Cytometry, Staining, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Immunohistochemistry

Journal: Acta biochimica et biophysica Sinica

Article Title: Modulation of ferroptosis via YY1-SLC7A11 axis in hepatic ischemia-reperfusion injury pathogenesis.

doi: 10.3724/abbs.2025093

Figure Lengend Snippet: Figure 3. Regulatory effect of NEDD4L expression in IRI on the ubiquitination-induced degradation of YY1 (A) Influence of MG-132 and leupeptin treatments on the reduction in YY1 expression induced by IRI-OGD/R, as detected by western blot analysis. (B) Detection of YY1 ubiquitination levels after IRI-OGD/R treatment. (C) Intersection of predicted human and mouse upstream E3 ligases of YY1, Venn diagram on the left, and the PPI network of 8 intersecting E3 ligases on the right (interaction score = 0.15). (D,E) Expression levels of 8 E3 ligases in mouse liver tissues and the AML12 cell line after IRI detected by RT-qPCR. (F) Immunohistochemical analysis of NEDD4L expression in mouse liver tissues after IRI-I/R treatment, representative images on the left, and quantitative statistical results on the right (scale bar = 50 μm). (G) Expressions of NEDD4L in mouse liver tissues and the AML12 cell line after IRI detected by western blot analysis. (H) Detection of YY1 expression and ubiquitination levels in AML12 cells overexpressing NEDD4L. (I) Co-IP experiment was used to detect the interaction between YY1 and NEDD4L in oe-NEDD4L. (J) Western blot analysis of the effects of NEDD4L knockdown on YY1 ubiquitination and expression after IRI-OGD/R treatment. (K) Flow cytometry analysis of the effects of NEDD4L knockdown on cell apoptosis induced by IRI-OGD/R; n = 10 per group in animal experiments. Cell experiments were repeated three times. *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: The AML12 normal mouse liver cell line (CRL2254; ATCC, Manassas, USA) was cultured in DMEM/F-12 (11320033; Thermo Fisher Scientific) supplemented with 10 μg/mL insulin, 5.5 μg/mL transferrin, 5 ng/mL selenium, 40 ng/mL dexamethasone, 10% FBS (10100147C; Thermo Fisher Scientific), and 1% penicillin-streptomycin (15140163; Thermo Fisher Scientific).

Techniques: Expressing, Ubiquitin Proteomics, Western Blot, Quantitative RT-PCR, Immunohistochemical staining, Co-Immunoprecipitation Assay, Knockdown, Flow Cytometry