cell lines hep 3b  (ATCC)

Journal: Scientific Reports

Article Title: Study on the effects and mechanisms of M2 macrophages on PYCR1-promoted biological behavior of hepatocellular carcinoma cells

doi: 10.1038/s41598-026-40817-8

Figure Lengend Snippet: Co-culture of M2 macrophage supernatant with hepatocellular carcinoma cells promotes HCC cell metastasis and EMT. ( A , B ) Wound healing assays of SNU-449 and Hep-3B cells under M0 and M2 supernatant co-culture conditions. Right panels show histogram results. Scale bar: 200 μm. ( C , D ) Transwell assays of SNU-449 and Hep-3B cells co-cultured with M0 and M2 supernatants. Scale bar: 50 μm. E Western blot analysis of EMT-related protein expression in SNU-449 and Hep-3B cells co-cultured with M0 and M2 supernatants ( n = 3 independent experiments). Data are presented as mean ± standard deviation (* P < 0.05, ** P < 0.01, *** P < 0.001 vs. Con group).

Article Snippet: The Hep-3B cell line of human hepatocellular carcinoma was obtained from Procell (Wuhan, China), while the SNU-449 cell line was acquired from the Chinese Academy of Sciences (Shanghai, China).

Techniques: Co-Culture Assay, Cell Culture, Western Blot, Expressing, Standard Deviation

Journal: Scientific Reports

Article Title: Study on the effects and mechanisms of M2 macrophages on PYCR1-promoted biological behavior of hepatocellular carcinoma cells

doi: 10.1038/s41598-026-40817-8

Figure Lengend Snippet: Knockdown of PYCR1 attenuates HCC cell metastasis and epithelial-mesenchymal transition induced by co-culture of M2 macrophage supernatant with hepatocellular carcinoma cells. ( A , B ) Wound healing assays of SNU-449 and Hep-3B cells with PYCR1 knockdown under altered co-culture conditions. Right panels show histograms of results. Scale bar: 200 μm. ( C , D ) Transwell assays assessing invasive migration of PYCR1-knockdown SNU-449 and Hep-3B cells under co-culture conditions. Scale bar: 50 μm. ( E , F ) Immunofluorescence Ki67 detection of the effect of PYCR1 knockdown on hepatocellular carcinoma cell proliferation under Co-culture Conditions ( n = 3 independent experiments). G Western blot analysis of EMT-related protein expression in PYCR1-knockdown SNU-449 and Hep-3B cells under co-culture conditions ( n = 3 independent experiments). (* P < 0.05, ** P < 0.01,*** P < 0.001, **** P < 0.0001 vs. NC + M0 group; ## P < 0.01, ### P < 0.001 vs. NC + M2 group)

Article Snippet: The Hep-3B cell line of human hepatocellular carcinoma was obtained from Procell (Wuhan, China), while the SNU-449 cell line was acquired from the Chinese Academy of Sciences (Shanghai, China).

Techniques: Knockdown, Co-Culture Assay, Migration, Immunofluorescence, Western Blot, Expressing

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: Real time monitoring of the autophagy process in untreated Hep3B cells. The cells were incubated for 48 h with complete growth medium. Hep3B cells were previously transfected with a plasmid expressing GFP-mRFP-LC3B. Green fluorescence is a marker of early autophagosomes. Instead, the red fluorescence remained stable after the fusion of the autophagosomes with lysosomes thus highlighting the late phase of autophagy.

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:

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: Real time monitoring of the autophagy process in Hep3B cells treated for 48 h with 2 mM oleic acid. Hep3B cells were previously transfected with a plasmid expressing GFP-mRFP-LC3B. Green fluorescence is a marker of early autophagosomes. Instead, the red fluorescence remained stable after the fusion of the autophagosomes with lysosomes thus highlighting the late phase of autophagy.

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:

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