human hepatoblastoma cell line hepg2  (ATCC)

Journal: Translational Gastroenterology and Hepatology

Article Title: Leptin-dependent fat accumulation triggers autophagy in metabolic dysfunction-associated steatohepatitis model

doi: 10.21037/tgh-25-17

Figure Lengend Snippet: Detection of the autophagy markers in liver cancer cells. (A) RT-qPCR detection of BECN1, MAP1LC3B, SQSTM1, UVRAG, TFEB, PRKAA1_1, and PRKAA2_1 in HepG2 cells after the administration of 2 mM OA, 100 pM BAF, 2 mM caffeine and their combination. The target expression was normalized to GAPDH. Data are presented as means ± SEM of treated vs. untreated cells from three independent experiments. (B,C) Western blot detection and densitometric quantification of the protein level of Beclin1, LC3B-I, LC3B-II, UVRAG, p62, AMPK-α and P-AMPK-α. Beta-actin was detected as equal loading control and used for the further densitometric normalization of the protein level of the target proteins. The error bars of the densitometric graph represent the SEM of experiments performed in triplicates. (D) Micrographs of MAP1LC3B-GFP-RFP stably transfected Hep3B cells after the administration of 2 mM OA (scale bar =300 µm). The cells were live monitored for up to 48 h (please refer to the ). *, P<0.05 by 2-way ANOVA with Dunnett’s test (see Appendix 1 for detailed statistical analysis). ANOVA, analysis of variance; OA, oleic acid; RT-qPCR, reverse transcription quantitative polymerase chain reaction; SEM, standard error mean.

Article Snippet: The human hepatoblastoma cell line HepG2 (ACC180, Leibniz Institute DSMZ-German Collection of Microorganisms and Cell Cultures GmbH, Braunschweig, Germany), the human hepatocellular carcinoma cell line Hep3B (ACC93, DSMZ) and the human HSC line LX-2, a kindly gift from Scott Friedmann (Icahn School of Medicine at Mount Sinai), were grown in DMEM (Gibco, Paisley, UK) supplemented with 10% fetal bovine serum, penicillin (100 U/mL) and streptomycin (100 μg/mL) (11548876, Thermo Fisher Scientific, Waltham, MA, USA) at 37 °C in a humidified atmosphere containing 5% CO 2 .

Techniques: Quantitative RT-PCR, Expressing, Western Blot, Control, Stable Transfection, Transfection, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: Translational Gastroenterology and Hepatology

Article Title: Leptin-dependent fat accumulation triggers autophagy in metabolic dysfunction-associated steatohepatitis model

doi: 10.21037/tgh-25-17

Figure Lengend Snippet: Autophagosome maturation monitoring in Hep3B treated with oleic acid. Intracellular lipid accumulation in HepG2 cells treated for 24 h with 500 ng/mL of rhTGF-β, 2 mM oleic acid, 100 pM bafilomycin, 2 mM caffeine and their combination. The lipids were stained with Oil Red O. Magnification: 100× (A,C) and 200× (B,D).

Article Snippet: The human hepatoblastoma cell line HepG2 (ACC180, Leibniz Institute DSMZ-German Collection of Microorganisms and Cell Cultures GmbH, Braunschweig, Germany), the human hepatocellular carcinoma cell line Hep3B (ACC93, DSMZ) and the human HSC line LX-2, a kindly gift from Scott Friedmann (Icahn School of Medicine at Mount Sinai), were grown in DMEM (Gibco, Paisley, UK) supplemented with 10% fetal bovine serum, penicillin (100 U/mL) and streptomycin (100 μg/mL) (11548876, Thermo Fisher Scientific, Waltham, MA, USA) at 37 °C in a humidified atmosphere containing 5% CO 2 .

Techniques: Staining