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human hepatocyte lo2 cells  (China Center for Type Culture Collection)

 
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    China Center for Type Culture Collection human hepatocyte lo2 cells
    Ferroptosis inhibitor attenuated ferroptosis-related death in <t>LO2</t> cells under PA treatment. (a) Representative Oil Red O staining of the LO2 cells cultured in 300 μ M PA for 24 h; scale bar: 100 μ m. (b) The quantitative analysis of Oil Red O–stained cells. (c) The viability of LO2 cells treated with PA or PA and Fer-1 was assessed. The cells were treated with PA (300 μ M) or PA (300 μ M) and Fer-1 (60 μ M) for 24 h, and then the viability of cells in each group was measured by the CCK-8 assay. (d) The level of the LDH released in LO2 cells treated with PA or PA and Fer-1 was measured using the LDH cytotoxicity assay kit. (e, f) The LO2 cells were stained with the LPO-specific dye BODIPY 581/591 C11, and LPO was detected by flow cytometry after the LO2 cells were treated with PA or PA and Fer-1. (g, h) LO2 cells were treated with PA or PA and Fer-1, stained with Annexin V-FITC and PI, and then cells that underwent apoptosis were quantified by flow cytometry; ⁣ ∗ p < 0.05, ⁣ ∗∗ p < 0.01, and ⁣ ∗∗∗ p < 0.001 between the indicated groups.
    Human Hepatocyte Lo2 Cells, supplied by China Center for Type Culture Collection, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/human+hepatocyte+lo2+cells/l02+cell+line/pmc11688137-74-0-8
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    Images

    1) Product Images from "Autophagy Regulates Ferroptosis-Mediated Diabetic Liver Injury by Modulating the Degradation of ACSL4"

    Article Title: Autophagy Regulates Ferroptosis-Mediated Diabetic Liver Injury by Modulating the Degradation of ACSL4

    Journal: Journal of Diabetes Research

    doi: 10.1155/jdr/7146054

    Ferroptosis inhibitor attenuated ferroptosis-related death in LO2 cells under PA treatment. (a) Representative Oil Red O staining of the LO2 cells cultured in 300 μ M PA for 24 h; scale bar: 100 μ m. (b) The quantitative analysis of Oil Red O–stained cells. (c) The viability of LO2 cells treated with PA or PA and Fer-1 was assessed. The cells were treated with PA (300 μ M) or PA (300 μ M) and Fer-1 (60 μ M) for 24 h, and then the viability of cells in each group was measured by the CCK-8 assay. (d) The level of the LDH released in LO2 cells treated with PA or PA and Fer-1 was measured using the LDH cytotoxicity assay kit. (e, f) The LO2 cells were stained with the LPO-specific dye BODIPY 581/591 C11, and LPO was detected by flow cytometry after the LO2 cells were treated with PA or PA and Fer-1. (g, h) LO2 cells were treated with PA or PA and Fer-1, stained with Annexin V-FITC and PI, and then cells that underwent apoptosis were quantified by flow cytometry; ⁣ ∗ p < 0.05, ⁣ ∗∗ p < 0.01, and ⁣ ∗∗∗ p < 0.001 between the indicated groups.
    Figure Legend Snippet: Ferroptosis inhibitor attenuated ferroptosis-related death in LO2 cells under PA treatment. (a) Representative Oil Red O staining of the LO2 cells cultured in 300 μ M PA for 24 h; scale bar: 100 μ m. (b) The quantitative analysis of Oil Red O–stained cells. (c) The viability of LO2 cells treated with PA or PA and Fer-1 was assessed. The cells were treated with PA (300 μ M) or PA (300 μ M) and Fer-1 (60 μ M) for 24 h, and then the viability of cells in each group was measured by the CCK-8 assay. (d) The level of the LDH released in LO2 cells treated with PA or PA and Fer-1 was measured using the LDH cytotoxicity assay kit. (e, f) The LO2 cells were stained with the LPO-specific dye BODIPY 581/591 C11, and LPO was detected by flow cytometry after the LO2 cells were treated with PA or PA and Fer-1. (g, h) LO2 cells were treated with PA or PA and Fer-1, stained with Annexin V-FITC and PI, and then cells that underwent apoptosis were quantified by flow cytometry; ⁣ ∗ p < 0.05, ⁣ ∗∗ p < 0.01, and ⁣ ∗∗∗ p < 0.001 between the indicated groups.

    Techniques Used: Staining, Cell Culture, CCK-8 Assay, LDH Cytotoxicity Assay, Flow Cytometry

    ACSL4 increased under PA treatment and was degraded through the autophagy-lysosomal pathway. (a) RT-qPCR analysis of the expression of ACSL4 mRNA in the LO2 cells treated with PA or PA and Fer-1. (b, c) Representative Western blots and quantitative densitometry analysis of ACSL4 protein expression in LO2 cells treated with PA or PA and Fer-1. (d, e) Protein stability of ACSL4 in LO2 cells cultured with PA, following a time course treatment with 100 μ g/mL CHX. (f, g) LO2 cells were treated with PA with or without MG132 (20 μ M) or Baf A1 (20 nM) for 8 h. Representative Western blots and quantitative densitometry analysis of the expression of the ACSL4 protein in LO2 cells. All data are presented as the mean ± Std Dev of at least three independent experiments; ⁣ ∗ p < 0.05 between the indicated groups.
    Figure Legend Snippet: ACSL4 increased under PA treatment and was degraded through the autophagy-lysosomal pathway. (a) RT-qPCR analysis of the expression of ACSL4 mRNA in the LO2 cells treated with PA or PA and Fer-1. (b, c) Representative Western blots and quantitative densitometry analysis of ACSL4 protein expression in LO2 cells treated with PA or PA and Fer-1. (d, e) Protein stability of ACSL4 in LO2 cells cultured with PA, following a time course treatment with 100 μ g/mL CHX. (f, g) LO2 cells were treated with PA with or without MG132 (20 μ M) or Baf A1 (20 nM) for 8 h. Representative Western blots and quantitative densitometry analysis of the expression of the ACSL4 protein in LO2 cells. All data are presented as the mean ± Std Dev of at least three independent experiments; ⁣ ∗ p < 0.05 between the indicated groups.

    Techniques Used: Quantitative RT-PCR, Expressing, Western Blot, Cell Culture

    Downregulation of ACSL4 reversed PA-induced ferroptosis in LO2 cells. (a) qPCR analysis of ACSL4 mRNA expression in PA300+siNC or PA300 + siACSL4 LO2 cells. (b) Immunoblotting analysis of ACSL4 protein expression in PA300+siNC or PA300+siACSL4 LO2 cells. (c) Cell viability of PA-stimulated LO2 cells transfected with control and siACSL4 knockdown RNA. (d) Rate of LPO of PA-stimulated LO2 cells transfected with control and siACSL4 knockdown RNA. Bars represent the mean ± Std Dev; ⁣ ∗∗ p < 0.01 between the indicated groups.
    Figure Legend Snippet: Downregulation of ACSL4 reversed PA-induced ferroptosis in LO2 cells. (a) qPCR analysis of ACSL4 mRNA expression in PA300+siNC or PA300 + siACSL4 LO2 cells. (b) Immunoblotting analysis of ACSL4 protein expression in PA300+siNC or PA300+siACSL4 LO2 cells. (c) Cell viability of PA-stimulated LO2 cells transfected with control and siACSL4 knockdown RNA. (d) Rate of LPO of PA-stimulated LO2 cells transfected with control and siACSL4 knockdown RNA. Bars represent the mean ± Std Dev; ⁣ ∗∗ p < 0.01 between the indicated groups.

    Techniques Used: Expressing, Western Blot, Transfection, Control, Knockdown

    Inhibition of autophagic flux in PA-treated LO2 cells. (a, b) Representative Western blots and quantitative densitometry analysis of the expression of LC3-II, Beclin1, and p62 proteins in the LO2 cells exposed to PA for 24 h. (c, d) The LO2 cells were treated with PA for 24 h in the presence or absence of 20 nM Baf A1 during the last 2 h of coculture. Representative Western blots and quantitative densitometry analysis of the expression of the LC3-II protein in LO2 cells. (e) Representative images of LO2 cells infected with mRFP-GFP-LC3 and exposed to PA; Green: GFP puncta; Red: mRFP puncta. (f) GFP dots and mRFP dots were counted in each cell. (g) Semiquantitative analysis of autophagosomes (yellow puncta in merged images) and autolysosomes (red puncta in merged images). All data are presented as the mean ± Std Dev of at least three separate experiments; ns: nonsignificant; ⁣ ∗ p < 0.05, ⁣ ∗∗ p < 0.01, and ⁣ ∗∗∗ p < 0.001 between the indicated groups.
    Figure Legend Snippet: Inhibition of autophagic flux in PA-treated LO2 cells. (a, b) Representative Western blots and quantitative densitometry analysis of the expression of LC3-II, Beclin1, and p62 proteins in the LO2 cells exposed to PA for 24 h. (c, d) The LO2 cells were treated with PA for 24 h in the presence or absence of 20 nM Baf A1 during the last 2 h of coculture. Representative Western blots and quantitative densitometry analysis of the expression of the LC3-II protein in LO2 cells. (e) Representative images of LO2 cells infected with mRFP-GFP-LC3 and exposed to PA; Green: GFP puncta; Red: mRFP puncta. (f) GFP dots and mRFP dots were counted in each cell. (g) Semiquantitative analysis of autophagosomes (yellow puncta in merged images) and autolysosomes (red puncta in merged images). All data are presented as the mean ± Std Dev of at least three separate experiments; ns: nonsignificant; ⁣ ∗ p < 0.05, ⁣ ∗∗ p < 0.01, and ⁣ ∗∗∗ p < 0.001 between the indicated groups.

    Techniques Used: Inhibition, Western Blot, Expressing, Infection

    Autophagy may be involved in ferroptosis by regulating the expression of ACSL4. (a) CCK-8 assay was performed to determine the viability of LO2 cells treated with various concentrations of 3-MA for 24 h. (b) The level of the LDH released in LO2 cells treated with various concentrations of 3-MA for 24 h was measured using the LDH cytotoxicity assay kit. (c, d) LO2 cells were treated with 5 mM 3-MA for 24 h and stained with the LPO-specific dye BODIPY 581/591 C11, and LPO was detected by flow cytometry. (e, f) LO2 cells were treated with 5 mM 3-MA for 24 h and stained with Annexin V-FITC and PI, and then the cells that underwent apoptosis were quantified by flow cytometry. (g, h) Representative Western blots and quantitative densitometry analysis of the expression of the ACSL4 and LC3-II proteins in LO2 cells cultured with 3-MA for 24 h. (i, j) LO2 cells were treated with BSA, BSA+Rap (50 nM), PA (300 μ M), or PA (300 μ M)+Rap (50 nM) for 24 h. Representative Western blots and quantitative densitometry analysis of the expression of ACSL4 and LC3 proteins in LO2 cells. All data are presented as the mean ± Std Dev of at least three separate experiments; ns: nonsignificant; ⁣ ∗ p < 0.05 and ⁣ ∗∗ p < 0.01 between the indicated groups.
    Figure Legend Snippet: Autophagy may be involved in ferroptosis by regulating the expression of ACSL4. (a) CCK-8 assay was performed to determine the viability of LO2 cells treated with various concentrations of 3-MA for 24 h. (b) The level of the LDH released in LO2 cells treated with various concentrations of 3-MA for 24 h was measured using the LDH cytotoxicity assay kit. (c, d) LO2 cells were treated with 5 mM 3-MA for 24 h and stained with the LPO-specific dye BODIPY 581/591 C11, and LPO was detected by flow cytometry. (e, f) LO2 cells were treated with 5 mM 3-MA for 24 h and stained with Annexin V-FITC and PI, and then the cells that underwent apoptosis were quantified by flow cytometry. (g, h) Representative Western blots and quantitative densitometry analysis of the expression of the ACSL4 and LC3-II proteins in LO2 cells cultured with 3-MA for 24 h. (i, j) LO2 cells were treated with BSA, BSA+Rap (50 nM), PA (300 μ M), or PA (300 μ M)+Rap (50 nM) for 24 h. Representative Western blots and quantitative densitometry analysis of the expression of ACSL4 and LC3 proteins in LO2 cells. All data are presented as the mean ± Std Dev of at least three separate experiments; ns: nonsignificant; ⁣ ∗ p < 0.05 and ⁣ ∗∗ p < 0.01 between the indicated groups.

    Techniques Used: Expressing, CCK-8 Assay, LDH Cytotoxicity Assay, Staining, Flow Cytometry, Western Blot, Cell Culture

    Related Articles

    Cell Culture:

    Article Title: Autophagy Regulates Ferroptosis-Mediated Diabetic Liver Injury by Modulating the Degradation of ACSL4
    Article Snippet: Ultrathin slices (100 nm) were observed using a JEM2000EX transmission electron microscope (JEOL, Tokyo, Japan) after being sliced and stained with lead citrate and uranyl acetate. .. Human hepatocyte LO2 cells were obtained from the China Center for Type Culture Collection (CCTCC, Wuhan, China) and cultured in Dulbecco's modified Eagle's medium (Gibco, United States) supplemented with 10% fetal bovine serum and 100 U/mL penicillin and streptomycin at 5% CO 2 . ..

    Modification:

    Article Title: Autophagy Regulates Ferroptosis-Mediated Diabetic Liver Injury by Modulating the Degradation of ACSL4
    Article Snippet: Ultrathin slices (100 nm) were observed using a JEM2000EX transmission electron microscope (JEOL, Tokyo, Japan) after being sliced and stained with lead citrate and uranyl acetate. .. Human hepatocyte LO2 cells were obtained from the China Center for Type Culture Collection (CCTCC, Wuhan, China) and cultured in Dulbecco's modified Eagle's medium (Gibco, United States) supplemented with 10% fetal bovine serum and 100 U/mL penicillin and streptomycin at 5% CO 2 . ..



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    Image Search Results


    Analysis of the expression of POC1A in various classifications of HCC patients, as well as in HCC and liver cancer cell lines. (A) Comparison of POC1A between cancer tissue samples and normal tissue samples. (B) Comparison of the expression of POC1A in different genders. (C) Comparison of the expression of POC1A in different age groups. (D) The expression levels of POC1A were all higher in the Caucasian, African-American, and Asian HCC patients than the normal patients. The expression of POC1A was significantly higher in LIHC tissue samples from Asian patients than LIHC tissue samples from Caucasian patients. (E) The expression of POC1A in individual cancer stages. (F) The expression of POC1A in different transistor grades. (G) The expression of POC1A in tumor (T) stage. (H) The expression of POC1A in node (N) stage. (I) The expression of POC1A in metastasis (M) stage. (J) Analysis of POC1A expression in normal liver tissue and cancer tissue samples from patients with hepatocellular carcinoma using immunohistochemical methods in the Human Protein Atlas database. Liver sample: https://www.proteinatlas.org/ENSG00000164087-POC1A/tissue/liver#img , hepatocellular samples: https://www.proteinatlas.org/ENSG00000164087-POC1A/pathology/liver+cancer#img . Scale bar: 100 µm. (K) Analysis of POC1A mRNA expression in LO2 and HepG2 cell lines by qPCR assay. *, P<0.05; **, P<0.01; -: no significance. TCGA, The Cancer Genome Atlas; HCC, hepatocellular carcinoma; LIHC, liver hepatocellular carcinoma; mRNA, messenger RNA; qPCR, quantitative polymerase chain reaction; POC1A , POC1 centriolar protein A.

    Journal: Translational Cancer Research

    Article Title: Analysis of the role of POC1A in the development and progression of hepatocellular carcinoma

    doi: 10.21037/tcr-23-2398

    Figure Lengend Snippet: Analysis of the expression of POC1A in various classifications of HCC patients, as well as in HCC and liver cancer cell lines. (A) Comparison of POC1A between cancer tissue samples and normal tissue samples. (B) Comparison of the expression of POC1A in different genders. (C) Comparison of the expression of POC1A in different age groups. (D) The expression levels of POC1A were all higher in the Caucasian, African-American, and Asian HCC patients than the normal patients. The expression of POC1A was significantly higher in LIHC tissue samples from Asian patients than LIHC tissue samples from Caucasian patients. (E) The expression of POC1A in individual cancer stages. (F) The expression of POC1A in different transistor grades. (G) The expression of POC1A in tumor (T) stage. (H) The expression of POC1A in node (N) stage. (I) The expression of POC1A in metastasis (M) stage. (J) Analysis of POC1A expression in normal liver tissue and cancer tissue samples from patients with hepatocellular carcinoma using immunohistochemical methods in the Human Protein Atlas database. Liver sample: https://www.proteinatlas.org/ENSG00000164087-POC1A/tissue/liver#img , hepatocellular samples: https://www.proteinatlas.org/ENSG00000164087-POC1A/pathology/liver+cancer#img . Scale bar: 100 µm. (K) Analysis of POC1A mRNA expression in LO2 and HepG2 cell lines by qPCR assay. *, P<0.05; **, P<0.01; -: no significance. TCGA, The Cancer Genome Atlas; HCC, hepatocellular carcinoma; LIHC, liver hepatocellular carcinoma; mRNA, messenger RNA; qPCR, quantitative polymerase chain reaction; POC1A , POC1 centriolar protein A.

    Article Snippet: The human normal hepatocyte cell line LO2 (Servicebio Technology Co., Ltd., Wuhan, China) was cultured in Roswell Park Memorial Institute 1640 medium, and the hepatoma cell line HepG2 (Procell Life Science & Technology Co., Ltd., Wuhan, China) was cultured in Dulbecco’s Modified Eagle Medium (Hyclone, Thermo Fisher Scientific Inc., Waltham, MA), supplemented with 10% fetal bovine serum and antibiotics (10,000 U/mL penicillin and 10 mg/mL streptomycin) (Procell Life Science & Technology Co., Ltd.).

    Techniques: Expressing, Comparison, Immunohistochemical staining, Real-time Polymerase Chain Reaction

    Ferroptosis inhibitor attenuated ferroptosis-related death in LO2 cells under PA treatment. (a) Representative Oil Red O staining of the LO2 cells cultured in 300 μ M PA for 24 h; scale bar: 100 μ m. (b) The quantitative analysis of Oil Red O–stained cells. (c) The viability of LO2 cells treated with PA or PA and Fer-1 was assessed. The cells were treated with PA (300 μ M) or PA (300 μ M) and Fer-1 (60 μ M) for 24 h, and then the viability of cells in each group was measured by the CCK-8 assay. (d) The level of the LDH released in LO2 cells treated with PA or PA and Fer-1 was measured using the LDH cytotoxicity assay kit. (e, f) The LO2 cells were stained with the LPO-specific dye BODIPY 581/591 C11, and LPO was detected by flow cytometry after the LO2 cells were treated with PA or PA and Fer-1. (g, h) LO2 cells were treated with PA or PA and Fer-1, stained with Annexin V-FITC and PI, and then cells that underwent apoptosis were quantified by flow cytometry; ⁣ ∗ p < 0.05, ⁣ ∗∗ p < 0.01, and ⁣ ∗∗∗ p < 0.001 between the indicated groups.

    Journal: Journal of Diabetes Research

    Article Title: Autophagy Regulates Ferroptosis-Mediated Diabetic Liver Injury by Modulating the Degradation of ACSL4

    doi: 10.1155/jdr/7146054

    Figure Lengend Snippet: Ferroptosis inhibitor attenuated ferroptosis-related death in LO2 cells under PA treatment. (a) Representative Oil Red O staining of the LO2 cells cultured in 300 μ M PA for 24 h; scale bar: 100 μ m. (b) The quantitative analysis of Oil Red O–stained cells. (c) The viability of LO2 cells treated with PA or PA and Fer-1 was assessed. The cells were treated with PA (300 μ M) or PA (300 μ M) and Fer-1 (60 μ M) for 24 h, and then the viability of cells in each group was measured by the CCK-8 assay. (d) The level of the LDH released in LO2 cells treated with PA or PA and Fer-1 was measured using the LDH cytotoxicity assay kit. (e, f) The LO2 cells were stained with the LPO-specific dye BODIPY 581/591 C11, and LPO was detected by flow cytometry after the LO2 cells were treated with PA or PA and Fer-1. (g, h) LO2 cells were treated with PA or PA and Fer-1, stained with Annexin V-FITC and PI, and then cells that underwent apoptosis were quantified by flow cytometry; ⁣ ∗ p < 0.05, ⁣ ∗∗ p < 0.01, and ⁣ ∗∗∗ p < 0.001 between the indicated groups.

    Article Snippet: Human hepatocyte LO2 cells were obtained from the China Center for Type Culture Collection (CCTCC, Wuhan, China) and cultured in Dulbecco's modified Eagle's medium (Gibco, United States) supplemented with 10% fetal bovine serum and 100 U/mL penicillin and streptomycin at 5% CO 2 .

    Techniques: Staining, Cell Culture, CCK-8 Assay, LDH Cytotoxicity Assay, Flow Cytometry

    ACSL4 increased under PA treatment and was degraded through the autophagy-lysosomal pathway. (a) RT-qPCR analysis of the expression of ACSL4 mRNA in the LO2 cells treated with PA or PA and Fer-1. (b, c) Representative Western blots and quantitative densitometry analysis of ACSL4 protein expression in LO2 cells treated with PA or PA and Fer-1. (d, e) Protein stability of ACSL4 in LO2 cells cultured with PA, following a time course treatment with 100 μ g/mL CHX. (f, g) LO2 cells were treated with PA with or without MG132 (20 μ M) or Baf A1 (20 nM) for 8 h. Representative Western blots and quantitative densitometry analysis of the expression of the ACSL4 protein in LO2 cells. All data are presented as the mean ± Std Dev of at least three independent experiments; ⁣ ∗ p < 0.05 between the indicated groups.

    Journal: Journal of Diabetes Research

    Article Title: Autophagy Regulates Ferroptosis-Mediated Diabetic Liver Injury by Modulating the Degradation of ACSL4

    doi: 10.1155/jdr/7146054

    Figure Lengend Snippet: ACSL4 increased under PA treatment and was degraded through the autophagy-lysosomal pathway. (a) RT-qPCR analysis of the expression of ACSL4 mRNA in the LO2 cells treated with PA or PA and Fer-1. (b, c) Representative Western blots and quantitative densitometry analysis of ACSL4 protein expression in LO2 cells treated with PA or PA and Fer-1. (d, e) Protein stability of ACSL4 in LO2 cells cultured with PA, following a time course treatment with 100 μ g/mL CHX. (f, g) LO2 cells were treated with PA with or without MG132 (20 μ M) or Baf A1 (20 nM) for 8 h. Representative Western blots and quantitative densitometry analysis of the expression of the ACSL4 protein in LO2 cells. All data are presented as the mean ± Std Dev of at least three independent experiments; ⁣ ∗ p < 0.05 between the indicated groups.

    Article Snippet: Human hepatocyte LO2 cells were obtained from the China Center for Type Culture Collection (CCTCC, Wuhan, China) and cultured in Dulbecco's modified Eagle's medium (Gibco, United States) supplemented with 10% fetal bovine serum and 100 U/mL penicillin and streptomycin at 5% CO 2 .

    Techniques: Quantitative RT-PCR, Expressing, Western Blot, Cell Culture

    Downregulation of ACSL4 reversed PA-induced ferroptosis in LO2 cells. (a) qPCR analysis of ACSL4 mRNA expression in PA300+siNC or PA300 + siACSL4 LO2 cells. (b) Immunoblotting analysis of ACSL4 protein expression in PA300+siNC or PA300+siACSL4 LO2 cells. (c) Cell viability of PA-stimulated LO2 cells transfected with control and siACSL4 knockdown RNA. (d) Rate of LPO of PA-stimulated LO2 cells transfected with control and siACSL4 knockdown RNA. Bars represent the mean ± Std Dev; ⁣ ∗∗ p < 0.01 between the indicated groups.

    Journal: Journal of Diabetes Research

    Article Title: Autophagy Regulates Ferroptosis-Mediated Diabetic Liver Injury by Modulating the Degradation of ACSL4

    doi: 10.1155/jdr/7146054

    Figure Lengend Snippet: Downregulation of ACSL4 reversed PA-induced ferroptosis in LO2 cells. (a) qPCR analysis of ACSL4 mRNA expression in PA300+siNC or PA300 + siACSL4 LO2 cells. (b) Immunoblotting analysis of ACSL4 protein expression in PA300+siNC or PA300+siACSL4 LO2 cells. (c) Cell viability of PA-stimulated LO2 cells transfected with control and siACSL4 knockdown RNA. (d) Rate of LPO of PA-stimulated LO2 cells transfected with control and siACSL4 knockdown RNA. Bars represent the mean ± Std Dev; ⁣ ∗∗ p < 0.01 between the indicated groups.

    Article Snippet: Human hepatocyte LO2 cells were obtained from the China Center for Type Culture Collection (CCTCC, Wuhan, China) and cultured in Dulbecco's modified Eagle's medium (Gibco, United States) supplemented with 10% fetal bovine serum and 100 U/mL penicillin and streptomycin at 5% CO 2 .

    Techniques: Expressing, Western Blot, Transfection, Control, Knockdown

    Inhibition of autophagic flux in PA-treated LO2 cells. (a, b) Representative Western blots and quantitative densitometry analysis of the expression of LC3-II, Beclin1, and p62 proteins in the LO2 cells exposed to PA for 24 h. (c, d) The LO2 cells were treated with PA for 24 h in the presence or absence of 20 nM Baf A1 during the last 2 h of coculture. Representative Western blots and quantitative densitometry analysis of the expression of the LC3-II protein in LO2 cells. (e) Representative images of LO2 cells infected with mRFP-GFP-LC3 and exposed to PA; Green: GFP puncta; Red: mRFP puncta. (f) GFP dots and mRFP dots were counted in each cell. (g) Semiquantitative analysis of autophagosomes (yellow puncta in merged images) and autolysosomes (red puncta in merged images). All data are presented as the mean ± Std Dev of at least three separate experiments; ns: nonsignificant; ⁣ ∗ p < 0.05, ⁣ ∗∗ p < 0.01, and ⁣ ∗∗∗ p < 0.001 between the indicated groups.

    Journal: Journal of Diabetes Research

    Article Title: Autophagy Regulates Ferroptosis-Mediated Diabetic Liver Injury by Modulating the Degradation of ACSL4

    doi: 10.1155/jdr/7146054

    Figure Lengend Snippet: Inhibition of autophagic flux in PA-treated LO2 cells. (a, b) Representative Western blots and quantitative densitometry analysis of the expression of LC3-II, Beclin1, and p62 proteins in the LO2 cells exposed to PA for 24 h. (c, d) The LO2 cells were treated with PA for 24 h in the presence or absence of 20 nM Baf A1 during the last 2 h of coculture. Representative Western blots and quantitative densitometry analysis of the expression of the LC3-II protein in LO2 cells. (e) Representative images of LO2 cells infected with mRFP-GFP-LC3 and exposed to PA; Green: GFP puncta; Red: mRFP puncta. (f) GFP dots and mRFP dots were counted in each cell. (g) Semiquantitative analysis of autophagosomes (yellow puncta in merged images) and autolysosomes (red puncta in merged images). All data are presented as the mean ± Std Dev of at least three separate experiments; ns: nonsignificant; ⁣ ∗ p < 0.05, ⁣ ∗∗ p < 0.01, and ⁣ ∗∗∗ p < 0.001 between the indicated groups.

    Article Snippet: Human hepatocyte LO2 cells were obtained from the China Center for Type Culture Collection (CCTCC, Wuhan, China) and cultured in Dulbecco's modified Eagle's medium (Gibco, United States) supplemented with 10% fetal bovine serum and 100 U/mL penicillin and streptomycin at 5% CO 2 .

    Techniques: Inhibition, Western Blot, Expressing, Infection

    Autophagy may be involved in ferroptosis by regulating the expression of ACSL4. (a) CCK-8 assay was performed to determine the viability of LO2 cells treated with various concentrations of 3-MA for 24 h. (b) The level of the LDH released in LO2 cells treated with various concentrations of 3-MA for 24 h was measured using the LDH cytotoxicity assay kit. (c, d) LO2 cells were treated with 5 mM 3-MA for 24 h and stained with the LPO-specific dye BODIPY 581/591 C11, and LPO was detected by flow cytometry. (e, f) LO2 cells were treated with 5 mM 3-MA for 24 h and stained with Annexin V-FITC and PI, and then the cells that underwent apoptosis were quantified by flow cytometry. (g, h) Representative Western blots and quantitative densitometry analysis of the expression of the ACSL4 and LC3-II proteins in LO2 cells cultured with 3-MA for 24 h. (i, j) LO2 cells were treated with BSA, BSA+Rap (50 nM), PA (300 μ M), or PA (300 μ M)+Rap (50 nM) for 24 h. Representative Western blots and quantitative densitometry analysis of the expression of ACSL4 and LC3 proteins in LO2 cells. All data are presented as the mean ± Std Dev of at least three separate experiments; ns: nonsignificant; ⁣ ∗ p < 0.05 and ⁣ ∗∗ p < 0.01 between the indicated groups.

    Journal: Journal of Diabetes Research

    Article Title: Autophagy Regulates Ferroptosis-Mediated Diabetic Liver Injury by Modulating the Degradation of ACSL4

    doi: 10.1155/jdr/7146054

    Figure Lengend Snippet: Autophagy may be involved in ferroptosis by regulating the expression of ACSL4. (a) CCK-8 assay was performed to determine the viability of LO2 cells treated with various concentrations of 3-MA for 24 h. (b) The level of the LDH released in LO2 cells treated with various concentrations of 3-MA for 24 h was measured using the LDH cytotoxicity assay kit. (c, d) LO2 cells were treated with 5 mM 3-MA for 24 h and stained with the LPO-specific dye BODIPY 581/591 C11, and LPO was detected by flow cytometry. (e, f) LO2 cells were treated with 5 mM 3-MA for 24 h and stained with Annexin V-FITC and PI, and then the cells that underwent apoptosis were quantified by flow cytometry. (g, h) Representative Western blots and quantitative densitometry analysis of the expression of the ACSL4 and LC3-II proteins in LO2 cells cultured with 3-MA for 24 h. (i, j) LO2 cells were treated with BSA, BSA+Rap (50 nM), PA (300 μ M), or PA (300 μ M)+Rap (50 nM) for 24 h. Representative Western blots and quantitative densitometry analysis of the expression of ACSL4 and LC3 proteins in LO2 cells. All data are presented as the mean ± Std Dev of at least three separate experiments; ns: nonsignificant; ⁣ ∗ p < 0.05 and ⁣ ∗∗ p < 0.01 between the indicated groups.

    Article Snippet: Human hepatocyte LO2 cells were obtained from the China Center for Type Culture Collection (CCTCC, Wuhan, China) and cultured in Dulbecco's modified Eagle's medium (Gibco, United States) supplemented with 10% fetal bovine serum and 100 U/mL penicillin and streptomycin at 5% CO 2 .

    Techniques: Expressing, CCK-8 Assay, LDH Cytotoxicity Assay, Staining, Flow Cytometry, Western Blot, Cell Culture