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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: 98/100, based on 1664 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/murine+normal+hepatocyte+aml12+cell+line/AML12/pmc12836525-191-1-13
    Average 98 stars, based on 1664 article reviews
    murine normal hepatocyte aml12 cell line - by Bioz Stars, 2026-10
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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

    Related Articles

    Control:

    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
    Article Snippet: Cell culture, transfection, and reagents: Human hepatocellular carcinoma cell lines without TM4SF5 expression (SNU449 and SNU761) and cell lines expressing TM4SF5 (Huh7, HepG2, and Hep3B) were acquired from the Korean Cell Line Bank (Seoul National University, Seoul, Korea).Cell culture, transfection, and reagents: Human hepatocellular carcinoma cell lines without TM4SF5 expression (SNU449 and SNU761) and cell lines expressing TM4SF5 (Huh7, HepG2, and Hep3B) were acquired from the Korean Cell Line Bank (Seoul National University, Seoul, Korea).. The murine normal hepatocyte AML12 cell line, lacking TM4SF5, was obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).. SNU449, SNU761, and Hep3B cell lines were cultured in RPMI-1640 medium (SH30027.01, Hyclone, Logan, UT, USA).SNU449, SNU761, and Hep3B cell lines were cultured in RPMI-1640 medium (SH30027.01, Hyclone, Logan, UT, USA).

    Stable Transfection:

    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
    Article Snippet: Cell culture, transfection, and reagents: Human hepatocellular carcinoma cell lines without TM4SF5 expression (SNU449 and SNU761) and cell lines expressing TM4SF5 (Huh7, HepG2, and Hep3B) were acquired from the Korean Cell Line Bank (Seoul National University, Seoul, Korea).Cell culture, transfection, and reagents: Human hepatocellular carcinoma cell lines without TM4SF5 expression (SNU449 and SNU761) and cell lines expressing TM4SF5 (Huh7, HepG2, and Hep3B) were acquired from the Korean Cell Line Bank (Seoul National University, Seoul, Korea).. The murine normal hepatocyte AML12 cell line, lacking TM4SF5, was obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).. SNU449, SNU761, and Hep3B cell lines were cultured in RPMI-1640 medium (SH30027.01, Hyclone, Logan, UT, USA).SNU449, SNU761, and Hep3B cell lines were cultured in RPMI-1640 medium (SH30027.01, Hyclone, Logan, UT, USA).

    Transfection:

    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
    Article Snippet: Cell culture, transfection, and reagents: Human hepatocellular carcinoma cell lines without TM4SF5 expression (SNU449 and SNU761) and cell lines expressing TM4SF5 (Huh7, HepG2, and Hep3B) were acquired from the Korean Cell Line Bank (Seoul National University, Seoul, Korea).Cell culture, transfection, and reagents: Human hepatocellular carcinoma cell lines without TM4SF5 expression (SNU449 and SNU761) and cell lines expressing TM4SF5 (Huh7, HepG2, and Hep3B) were acquired from the Korean Cell Line Bank (Seoul National University, Seoul, Korea).. The murine normal hepatocyte AML12 cell line, lacking TM4SF5, was obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).. SNU449, SNU761, and Hep3B cell lines were cultured in RPMI-1640 medium (SH30027.01, Hyclone, Logan, UT, USA).SNU449, SNU761, and Hep3B cell lines were cultured in RPMI-1640 medium (SH30027.01, Hyclone, Logan, UT, USA).

    Flow Cytometry:

    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
    Article Snippet: Cell culture, transfection, and reagents: Human hepatocellular carcinoma cell lines without TM4SF5 expression (SNU449 and SNU761) and cell lines expressing TM4SF5 (Huh7, HepG2, and Hep3B) were acquired from the Korean Cell Line Bank (Seoul National University, Seoul, Korea).Cell culture, transfection, and reagents: Human hepatocellular carcinoma cell lines without TM4SF5 expression (SNU449 and SNU761) and cell lines expressing TM4SF5 (Huh7, HepG2, and Hep3B) were acquired from the Korean Cell Line Bank (Seoul National University, Seoul, Korea).. The murine normal hepatocyte AML12 cell line, lacking TM4SF5, was obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).. SNU449, SNU761, and Hep3B cell lines were cultured in RPMI-1640 medium (SH30027.01, Hyclone, Logan, UT, USA).SNU449, SNU761, and Hep3B cell lines were cultured in RPMI-1640 medium (SH30027.01, Hyclone, Logan, UT, USA).

    Staining:

    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
    Article Snippet: Cell culture, transfection, and reagents: Human hepatocellular carcinoma cell lines without TM4SF5 expression (SNU449 and SNU761) and cell lines expressing TM4SF5 (Huh7, HepG2, and Hep3B) were acquired from the Korean Cell Line Bank (Seoul National University, Seoul, Korea).Cell culture, transfection, and reagents: Human hepatocellular carcinoma cell lines without TM4SF5 expression (SNU449 and SNU761) and cell lines expressing TM4SF5 (Huh7, HepG2, and Hep3B) were acquired from the Korean Cell Line Bank (Seoul National University, Seoul, Korea).. The murine normal hepatocyte AML12 cell line, lacking TM4SF5, was obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).. SNU449, SNU761, and Hep3B cell lines were cultured in RPMI-1640 medium (SH30027.01, Hyclone, Logan, UT, USA).SNU449, SNU761, and Hep3B cell lines were cultured in RPMI-1640 medium (SH30027.01, Hyclone, Logan, UT, USA).

    Knockdown:

    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
    Article Snippet: Cell culture, transfection, and reagents: Human hepatocellular carcinoma cell lines without TM4SF5 expression (SNU449 and SNU761) and cell lines expressing TM4SF5 (Huh7, HepG2, and Hep3B) were acquired from the Korean Cell Line Bank (Seoul National University, Seoul, Korea).Cell culture, transfection, and reagents: Human hepatocellular carcinoma cell lines without TM4SF5 expression (SNU449 and SNU761) and cell lines expressing TM4SF5 (Huh7, HepG2, and Hep3B) were acquired from the Korean Cell Line Bank (Seoul National University, Seoul, Korea).. The murine normal hepatocyte AML12 cell line, lacking TM4SF5, was obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).. SNU449, SNU761, and Hep3B cell lines were cultured in RPMI-1640 medium (SH30027.01, Hyclone, Logan, UT, USA).SNU449, SNU761, and Hep3B cell lines were cultured in RPMI-1640 medium (SH30027.01, Hyclone, Logan, UT, USA).

    Sequencing:

    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
    Article Snippet: Cell culture, transfection, and reagents: Human hepatocellular carcinoma cell lines without TM4SF5 expression (SNU449 and SNU761) and cell lines expressing TM4SF5 (Huh7, HepG2, and Hep3B) were acquired from the Korean Cell Line Bank (Seoul National University, Seoul, Korea).Cell culture, transfection, and reagents: Human hepatocellular carcinoma cell lines without TM4SF5 expression (SNU449 and SNU761) and cell lines expressing TM4SF5 (Huh7, HepG2, and Hep3B) were acquired from the Korean Cell Line Bank (Seoul National University, Seoul, Korea).. The murine normal hepatocyte AML12 cell line, lacking TM4SF5, was obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).. SNU449, SNU761, and Hep3B cell lines were cultured in RPMI-1640 medium (SH30027.01, Hyclone, Logan, UT, USA).SNU449, SNU761, and Hep3B cell lines were cultured in RPMI-1640 medium (SH30027.01, Hyclone, Logan, UT, USA).

    Expressing:

    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
    Article Snippet: Cell culture, transfection, and reagents: Human hepatocellular carcinoma cell lines without TM4SF5 expression (SNU449 and SNU761) and cell lines expressing TM4SF5 (Huh7, HepG2, and Hep3B) were acquired from the Korean Cell Line Bank (Seoul National University, Seoul, Korea).Cell culture, transfection, and reagents: Human hepatocellular carcinoma cell lines without TM4SF5 expression (SNU449 and SNU761) and cell lines expressing TM4SF5 (Huh7, HepG2, and Hep3B) were acquired from the Korean Cell Line Bank (Seoul National University, Seoul, Korea).. The murine normal hepatocyte AML12 cell line, lacking TM4SF5, was obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).. SNU449, SNU761, and Hep3B cell lines were cultured in RPMI-1640 medium (SH30027.01, Hyclone, Logan, UT, USA).SNU449, SNU761, and Hep3B cell lines were cultured in RPMI-1640 medium (SH30027.01, Hyclone, Logan, UT, USA).

    Plasmid Preparation:

    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
    Article Snippet: Cell culture, transfection, and reagents: Human hepatocellular carcinoma cell lines without TM4SF5 expression (SNU449 and SNU761) and cell lines expressing TM4SF5 (Huh7, HepG2, and Hep3B) were acquired from the Korean Cell Line Bank (Seoul National University, Seoul, Korea).Cell culture, transfection, and reagents: Human hepatocellular carcinoma cell lines without TM4SF5 expression (SNU449 and SNU761) and cell lines expressing TM4SF5 (Huh7, HepG2, and Hep3B) were acquired from the Korean Cell Line Bank (Seoul National University, Seoul, Korea).. The murine normal hepatocyte AML12 cell line, lacking TM4SF5, was obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).. SNU449, SNU761, and Hep3B cell lines were cultured in RPMI-1640 medium (SH30027.01, Hyclone, Logan, UT, USA).SNU449, SNU761, and Hep3B cell lines were cultured in RPMI-1640 medium (SH30027.01, Hyclone, Logan, UT, USA).

    Quantitative RT-PCR:

    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
    Article Snippet: Cell culture, transfection, and reagents: Human hepatocellular carcinoma cell lines without TM4SF5 expression (SNU449 and SNU761) and cell lines expressing TM4SF5 (Huh7, HepG2, and Hep3B) were acquired from the Korean Cell Line Bank (Seoul National University, Seoul, Korea).Cell culture, transfection, and reagents: Human hepatocellular carcinoma cell lines without TM4SF5 expression (SNU449 and SNU761) and cell lines expressing TM4SF5 (Huh7, HepG2, and Hep3B) were acquired from the Korean Cell Line Bank (Seoul National University, Seoul, Korea).. The murine normal hepatocyte AML12 cell line, lacking TM4SF5, was obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).. SNU449, SNU761, and Hep3B cell lines were cultured in RPMI-1640 medium (SH30027.01, Hyclone, Logan, UT, USA).SNU449, SNU761, and Hep3B cell lines were cultured in RPMI-1640 medium (SH30027.01, Hyclone, Logan, UT, USA).

    Western Blot:

    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
    Article Snippet: Cell culture, transfection, and reagents: Human hepatocellular carcinoma cell lines without TM4SF5 expression (SNU449 and SNU761) and cell lines expressing TM4SF5 (Huh7, HepG2, and Hep3B) were acquired from the Korean Cell Line Bank (Seoul National University, Seoul, Korea).Cell culture, transfection, and reagents: Human hepatocellular carcinoma cell lines without TM4SF5 expression (SNU449 and SNU761) and cell lines expressing TM4SF5 (Huh7, HepG2, and Hep3B) were acquired from the Korean Cell Line Bank (Seoul National University, Seoul, Korea).. The murine normal hepatocyte AML12 cell line, lacking TM4SF5, was obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).. SNU449, SNU761, and Hep3B cell lines were cultured in RPMI-1640 medium (SH30027.01, Hyclone, Logan, UT, USA).SNU449, SNU761, and Hep3B cell lines were cultured in RPMI-1640 medium (SH30027.01, Hyclone, Logan, UT, USA).



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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