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murine normal hepatocyte aml12 cell line  (ATCC)


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    ATCC murine normal hepatocyte aml12 cell line
    TM4SF5-mediated stabilization of KEAP1 in lipid-treated hepatocytes promotes oxidative stress and hepatic inflammation. (A-F) Subconfluent Huh7 Control or Huh7 TM4SF5-KO hepatocyte variants, either stably or transiently transfected with the indicated cDNAs, were analyzed for ROS production using flow cytometry following DCFDA staining. Cells received vehicle treatment (-), PA alone (B, D, and F), lipid mixture (LM, C), or DOX to induce Keap1 knockdown (shKEAP1 #2 or #3 , see Table , C and D). In some experiments, Huh7 cell variants were transfected with siNS (non-specific sequences as a control) or siNRF2 (targeting sequence #1 or #2, see Table ) for 24 h. Cells were then treated with DOX for shKEAP1 #2 induction for 24 h, followed by PA for 4 h prior to ROS analysis by flow cytometry (D). Cells were also exposed to H 2 O 2 at the indicated concentrations and durations before assessment of ROS levels (E). *, P < 0.05; **, P < 0.01; ***, P < 0.001; ***, P < 0.0001; ns = non-significant, unpaired Student's t test or two-way ANOVA. Data are expressed as mean ± SEM. (G) Subconfluent SNU449 hepatocytes stably expressing empty vector (SNU449 EV ) or TM4SF5 (SNU449 TM4SF5 ) were harvested to perform qRT-PCR for the indicated molecules. (H) Subconfluent Huh7 Control cells were treated in the absence (-) or presence (+) of DOX to induce KEAP1 knockdown (shKEAP1 #2 ) for 24 h and then exposed to PA (100 μM) for 4 h before being collected for immunoblot analysis of the indicated molecules. (I) Murine <t>AML12</t> cells stably transfected with EV or TM4SF5 were treated with 5% LM for 24 h, harvested, and subjected to qRT-PCR for the indicated molecules. Data shown are representative of three independent experiments. See also .
    Murine Normal Hepatocyte Aml12 Cell Line, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1687 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "TM4SF5-mediated KEAP1 Regulation in Hepatocytes Irrelevant to NRF2 Expression and Activity Promotes Oxidative Stress and Inflammation to Develop Metabolic Dysfunction-Associated Steatotic Liver Disease"

    Article Title: TM4SF5-mediated KEAP1 Regulation in Hepatocytes Irrelevant to NRF2 Expression and Activity Promotes Oxidative Stress and Inflammation to Develop Metabolic Dysfunction-Associated Steatotic Liver Disease

    Journal: International Journal of Biological Sciences

    doi: 10.7150/ijbs.126251

    TM4SF5-mediated stabilization of KEAP1 in lipid-treated hepatocytes promotes oxidative stress and hepatic inflammation. (A-F) Subconfluent Huh7 Control or Huh7 TM4SF5-KO hepatocyte variants, either stably or transiently transfected with the indicated cDNAs, were analyzed for ROS production using flow cytometry following DCFDA staining. Cells received vehicle treatment (-), PA alone (B, D, and F), lipid mixture (LM, C), or DOX to induce Keap1 knockdown (shKEAP1 #2 or #3 , see Table , C and D). In some experiments, Huh7 cell variants were transfected with siNS (non-specific sequences as a control) or siNRF2 (targeting sequence #1 or #2, see Table ) for 24 h. Cells were then treated with DOX for shKEAP1 #2 induction for 24 h, followed by PA for 4 h prior to ROS analysis by flow cytometry (D). Cells were also exposed to H 2 O 2 at the indicated concentrations and durations before assessment of ROS levels (E). *, P < 0.05; **, P < 0.01; ***, P < 0.001; ***, P < 0.0001; ns = non-significant, unpaired Student's t test or two-way ANOVA. Data are expressed as mean ± SEM. (G) Subconfluent SNU449 hepatocytes stably expressing empty vector (SNU449 EV ) or TM4SF5 (SNU449 TM4SF5 ) were harvested to perform qRT-PCR for the indicated molecules. (H) Subconfluent Huh7 Control cells were treated in the absence (-) or presence (+) of DOX to induce KEAP1 knockdown (shKEAP1 #2 ) for 24 h and then exposed to PA (100 μM) for 4 h before being collected for immunoblot analysis of the indicated molecules. (I) Murine AML12 cells stably transfected with EV or TM4SF5 were treated with 5% LM for 24 h, harvested, and subjected to qRT-PCR for the indicated molecules. Data shown are representative of three independent experiments. See also .
    Figure Legend Snippet: TM4SF5-mediated stabilization of KEAP1 in lipid-treated hepatocytes promotes oxidative stress and hepatic inflammation. (A-F) Subconfluent Huh7 Control or Huh7 TM4SF5-KO hepatocyte variants, either stably or transiently transfected with the indicated cDNAs, were analyzed for ROS production using flow cytometry following DCFDA staining. Cells received vehicle treatment (-), PA alone (B, D, and F), lipid mixture (LM, C), or DOX to induce Keap1 knockdown (shKEAP1 #2 or #3 , see Table , C and D). In some experiments, Huh7 cell variants were transfected with siNS (non-specific sequences as a control) or siNRF2 (targeting sequence #1 or #2, see Table ) for 24 h. Cells were then treated with DOX for shKEAP1 #2 induction for 24 h, followed by PA for 4 h prior to ROS analysis by flow cytometry (D). Cells were also exposed to H 2 O 2 at the indicated concentrations and durations before assessment of ROS levels (E). *, P < 0.05; **, P < 0.01; ***, P < 0.001; ***, P < 0.0001; ns = non-significant, unpaired Student's t test or two-way ANOVA. Data are expressed as mean ± SEM. (G) Subconfluent SNU449 hepatocytes stably expressing empty vector (SNU449 EV ) or TM4SF5 (SNU449 TM4SF5 ) were harvested to perform qRT-PCR for the indicated molecules. (H) Subconfluent Huh7 Control cells were treated in the absence (-) or presence (+) of DOX to induce KEAP1 knockdown (shKEAP1 #2 ) for 24 h and then exposed to PA (100 μM) for 4 h before being collected for immunoblot analysis of the indicated molecules. (I) Murine AML12 cells stably transfected with EV or TM4SF5 were treated with 5% LM for 24 h, harvested, and subjected to qRT-PCR for the indicated molecules. Data shown are representative of three independent experiments. See also .

    Techniques Used: Control, Stable Transfection, Transfection, Flow Cytometry, Staining, Knockdown, Sequencing, Expressing, Plasmid Preparation, Quantitative RT-PCR, Western Blot



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    ATCC murine normal hepatocyte aml12 cell line
    TM4SF5-mediated stabilization of KEAP1 in lipid-treated hepatocytes promotes oxidative stress and hepatic inflammation. (A-F) Subconfluent Huh7 Control or Huh7 TM4SF5-KO hepatocyte variants, either stably or transiently transfected with the indicated cDNAs, were analyzed for ROS production using flow cytometry following DCFDA staining. Cells received vehicle treatment (-), PA alone (B, D, and F), lipid mixture (LM, C), or DOX to induce Keap1 knockdown (shKEAP1 #2 or #3 , see Table , C and D). In some experiments, Huh7 cell variants were transfected with siNS (non-specific sequences as a control) or siNRF2 (targeting sequence #1 or #2, see Table ) for 24 h. Cells were then treated with DOX for shKEAP1 #2 induction for 24 h, followed by PA for 4 h prior to ROS analysis by flow cytometry (D). Cells were also exposed to H 2 O 2 at the indicated concentrations and durations before assessment of ROS levels (E). *, P < 0.05; **, P < 0.01; ***, P < 0.001; ***, P < 0.0001; ns = non-significant, unpaired Student's t test or two-way ANOVA. Data are expressed as mean ± SEM. (G) Subconfluent SNU449 hepatocytes stably expressing empty vector (SNU449 EV ) or TM4SF5 (SNU449 TM4SF5 ) were harvested to perform qRT-PCR for the indicated molecules. (H) Subconfluent Huh7 Control cells were treated in the absence (-) or presence (+) of DOX to induce KEAP1 knockdown (shKEAP1 #2 ) for 24 h and then exposed to PA (100 μM) for 4 h before being collected for immunoblot analysis of the indicated molecules. (I) Murine <t>AML12</t> cells stably transfected with EV or TM4SF5 were treated with 5% LM for 24 h, harvested, and subjected to qRT-PCR for the indicated molecules. Data shown are representative of three independent experiments. See also .
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    TM4SF5-mediated stabilization of KEAP1 in lipid-treated hepatocytes promotes oxidative stress and hepatic inflammation. (A-F) Subconfluent Huh7 Control or Huh7 TM4SF5-KO hepatocyte variants, either stably or transiently transfected with the indicated cDNAs, were analyzed for ROS production using flow cytometry following DCFDA staining. Cells received vehicle treatment (-), PA alone (B, D, and F), lipid mixture (LM, C), or DOX to induce Keap1 knockdown (shKEAP1 #2 or #3 , see Table , C and D). In some experiments, Huh7 cell variants were transfected with siNS (non-specific sequences as a control) or siNRF2 (targeting sequence #1 or #2, see Table ) for 24 h. Cells were then treated with DOX for shKEAP1 #2 induction for 24 h, followed by PA for 4 h prior to ROS analysis by flow cytometry (D). Cells were also exposed to H 2 O 2 at the indicated concentrations and durations before assessment of ROS levels (E). *, P < 0.05; **, P < 0.01; ***, P < 0.001; ***, P < 0.0001; ns = non-significant, unpaired Student's t test or two-way ANOVA. Data are expressed as mean ± SEM. (G) Subconfluent SNU449 hepatocytes stably expressing empty vector (SNU449 EV ) or TM4SF5 (SNU449 TM4SF5 ) were harvested to perform qRT-PCR for the indicated molecules. (H) Subconfluent Huh7 Control cells were treated in the absence (-) or presence (+) of DOX to induce KEAP1 knockdown (shKEAP1 #2 ) for 24 h and then exposed to PA (100 μM) for 4 h before being collected for immunoblot analysis of the indicated molecules. (I) Murine AML12 cells stably transfected with EV or TM4SF5 were treated with 5% LM for 24 h, harvested, and subjected to qRT-PCR for the indicated molecules. Data shown are representative of three independent experiments. See also .

    Journal: International Journal of Biological Sciences

    Article Title: TM4SF5-mediated KEAP1 Regulation in Hepatocytes Irrelevant to NRF2 Expression and Activity Promotes Oxidative Stress and Inflammation to Develop Metabolic Dysfunction-Associated Steatotic Liver Disease

    doi: 10.7150/ijbs.126251

    Figure Lengend Snippet: TM4SF5-mediated stabilization of KEAP1 in lipid-treated hepatocytes promotes oxidative stress and hepatic inflammation. (A-F) Subconfluent Huh7 Control or Huh7 TM4SF5-KO hepatocyte variants, either stably or transiently transfected with the indicated cDNAs, were analyzed for ROS production using flow cytometry following DCFDA staining. Cells received vehicle treatment (-), PA alone (B, D, and F), lipid mixture (LM, C), or DOX to induce Keap1 knockdown (shKEAP1 #2 or #3 , see Table , C and D). In some experiments, Huh7 cell variants were transfected with siNS (non-specific sequences as a control) or siNRF2 (targeting sequence #1 or #2, see Table ) for 24 h. Cells were then treated with DOX for shKEAP1 #2 induction for 24 h, followed by PA for 4 h prior to ROS analysis by flow cytometry (D). Cells were also exposed to H 2 O 2 at the indicated concentrations and durations before assessment of ROS levels (E). *, P < 0.05; **, P < 0.01; ***, P < 0.001; ***, P < 0.0001; ns = non-significant, unpaired Student's t test or two-way ANOVA. Data are expressed as mean ± SEM. (G) Subconfluent SNU449 hepatocytes stably expressing empty vector (SNU449 EV ) or TM4SF5 (SNU449 TM4SF5 ) were harvested to perform qRT-PCR for the indicated molecules. (H) Subconfluent Huh7 Control cells were treated in the absence (-) or presence (+) of DOX to induce KEAP1 knockdown (shKEAP1 #2 ) for 24 h and then exposed to PA (100 μM) for 4 h before being collected for immunoblot analysis of the indicated molecules. (I) Murine AML12 cells stably transfected with EV or TM4SF5 were treated with 5% LM for 24 h, harvested, and subjected to qRT-PCR for the indicated molecules. Data shown are representative of three independent experiments. See also .

    Article Snippet: The murine normal hepatocyte AML12 cell line, lacking TM4SF5, was obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).

    Techniques: Control, Stable Transfection, Transfection, Flow Cytometry, Staining, Knockdown, Sequencing, Expressing, Plasmid Preparation, Quantitative RT-PCR, Western Blot

    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

    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:

    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)

    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