Journal: Antioxidants

Article Title: NRF2 Activation Ameliorates Oxidative Stress and Improves Mitochondrial Function and Synaptic Plasticity, and in A53T α-Synuclein Hippocampal Neurons

doi: 10.3390/antiox11010026

Figure Lengend Snippet: DMF activates NRF2 in vitro and this activation is inhibited by ML385. ( A ) 20 µM DMF significantly induces NRF2 expression in HepG2-ARE cells, and co-treatment with 1 µM ML385 significantly inhibits this effect. ( B ) 20 µM DMF significantly induced expression of NRF2-ARE genes, GCLC, HMOX1, and NQO1 in WT and A53TSyn neurons ( n = 10–12). Significance is relative to control unless otherwise indicated ( * p < 0.05, ** p < 0.01, *** p < 0.001).

Article Snippet: HepG2 cells that stably express a firefly luciferase gene under the control of the ARE promoter were obtained from BPS Bioscience.

Techniques: In Vitro, Activation Assay, Expressing

Journal: Hepatitis Monthly

Article Title: Activation of Nrf2-Antioxidant Response Element Mediated Glutamate Cysteine Ligase Expression in Hepatoma Cell line by Homocysteine

doi: 10.5812/hepatmon.8394

Figure Lengend Snippet: HepG2 cells were treated with D/L homocysteine (50 µM) for the indicated times, and harvested for flow cytometric analysis of intracellular glutathione content using mean channel fluorescence (MCF) of ThiolTracker Violet dye (TTV). Analysis of cellular GSH content was restricted to PI negative intact cells. Values are means ± SE. Statistically different from control.

Article Snippet: The HepG2 cells (National Cell Bank, Pasteur Institute of Iran) were cultured in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10 % fetal calf serum and 100 u/ml penicillin G sodium, 100 μg/ml streptomycinand L-glutamine in humidified atmosphere in 5% CO2 at 37oC.

Techniques: Fluorescence, Control

Journal: Hepatitis Monthly

Article Title: Activation of Nrf2-Antioxidant Response Element Mediated Glutamate Cysteine Ligase Expression in Hepatoma Cell line by Homocysteine

doi: 10.5812/hepatmon.8394

Figure Lengend Snippet: HepG2 cells were treated with D/L homocysteine (50 µM) for the indicated time points and GCLc mRNA levels were determined using quantitative real-time RT-PCR analysis. The bar graph shows the quantization of GCLc gene expression. Values are normalized to β-actin expression and represent the means ± SE of four separate experiments.

Article Snippet: The HepG2 cells (National Cell Bank, Pasteur Institute of Iran) were cultured in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10 % fetal calf serum and 100 u/ml penicillin G sodium, 100 μg/ml streptomycinand L-glutamine in humidified atmosphere in 5% CO2 at 37oC.

Techniques: Quantitative RT-PCR, Gene Expression, Expressing

Journal: Hepatitis Monthly

Article Title: Activation of Nrf2-Antioxidant Response Element Mediated Glutamate Cysteine Ligase Expression in Hepatoma Cell line by Homocysteine

doi: 10.5812/hepatmon.8394

Figure Lengend Snippet: A) The protein levels were detected by western blotting. Values represent the means ± SE of three separate experiments. The bar graph shows the quantization of GCLc protein B) Western blot analysis of GCLc in HepG2 cells. The cells were incubated with 50 μM homocysteine for 3, 6 and 9 h, and the cell extracts were electrophoresed, protein transferred, and blotted with the polyclonal antibody to GCLc. GCLc is a major band appearing at approximately 73KD. β-actin was used as loading control.

Article Snippet: The HepG2 cells (National Cell Bank, Pasteur Institute of Iran) were cultured in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10 % fetal calf serum and 100 u/ml penicillin G sodium, 100 μg/ml streptomycinand L-glutamine in humidified atmosphere in 5% CO2 at 37oC.

Techniques: Western Blot, Incubation, Control

Journal: Hepatitis Monthly

Article Title: Activation of Nrf2-Antioxidant Response Element Mediated Glutamate Cysteine Ligase Expression in Hepatoma Cell line by Homocysteine

doi: 10.5812/hepatmon.8394

Figure Lengend Snippet: A) Nrf2 protein levels in nuclear fractions of HepG2 cells exposed to homocysteine (50 µM) for the indicated time. The nuclear cell extracts were electrophoresed, protein transferred, and blotted with the polyclonal antibody Nrf2. Nrf2 is a major band appearing at approximately 68 kD. Lamin B was used as loading control. Quantification of band intensity was performed by Image J (version 1.46a). The bar graph shows the quantization of Nrf2 protein. Values represent the means ± SE of three separate experiments B) Gel shift assay using oligomers containing the GCLc promoter-specific ARE-biding site and nuclear fractions of HepG2 cells that were exposed to homocysteine for the indicated time periods. Competitor (200 fold excess) and Nrf2 antibody were applied as indicated.

Article Snippet: The HepG2 cells (National Cell Bank, Pasteur Institute of Iran) were cultured in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10 % fetal calf serum and 100 u/ml penicillin G sodium, 100 μg/ml streptomycinand L-glutamine in humidified atmosphere in 5% CO2 at 37oC.

Techniques: Control, Gel Shift

Journal: Molecules

Article Title: Simplified LC-MS Method for Analysis of Sterols in Biological Samples

doi: 10.3390/molecules25184116

Figure Lengend Snippet: Representative chromatograms of sterols isolated from native HepG2 cells ( a , b ) and HepG2 DHCR KO cells ( c , d ). Each chromatogram is presented in two magnifications to account for a higher quantity of cholesterol compared to intermediate sterols ( a , b ) or desmosterol and cholesterol compared to other sterols ( c , d ). Zymosterol (A), 24-dehydrolathosterol (B), 7-dehydrodesmosterol (C), 7-desmosterol (D), zymostenol (E), F-lathosterol (F), lathosterol-d7 (G), TMAS (I), holesterol (J), lanosterol (K), dihydrolanosterol (L), and an unknown peak with the MRM of 365/199 (X). All peaks with the same MRM transition (A, B, D and E, F, J) are chromatographically separated. Peak of cholesterol is depicted for illustrative purpose only.

Article Snippet: Here, 1 × 10 6 of HepG2 and HepG2 with DHCR24 deletion (ordered from Synthego, Menalo Park, CA, USA) cells were plated on a T75 culture flask for each sterol isolation.

Techniques: Isolation

Journal: Molecules

Article Title: Simplified LC-MS Method for Analysis of Sterols in Biological Samples

doi: 10.3390/molecules25184116

Figure Lengend Snippet: Concentrations of sterol intermediates isolated from HepG2 and HepG2 DHCR-KO cell lines. All concentrations are calculated as ng/mL per 10 7 , to normalize sterol concentration based on the number of counted cells. All data are represented as mean with SD, black dots represent individual measurements.

Article Snippet: Here, 1 × 10 6 of HepG2 and HepG2 with DHCR24 deletion (ordered from Synthego, Menalo Park, CA, USA) cells were plated on a T75 culture flask for each sterol isolation.

Techniques: Isolation, Concentration Assay

Journal: Molecules

Article Title: Simplified LC-MS Method for Analysis of Sterols in Biological Samples

doi: 10.3390/molecules25184116

Figure Lengend Snippet: Concentration of sterols isolated from HepG2 and HepG2 DHCR24 KO cell cultures.

Article Snippet: Here, 1 × 10 6 of HepG2 and HepG2 with DHCR24 deletion (ordered from Synthego, Menalo Park, CA, USA) cells were plated on a T75 culture flask for each sterol isolation.

Techniques: Concentration Assay, Isolation

Journal: Scientific Reports

Article Title: α-Lipoic acid induces Endoplasmic Reticulum stress-mediated apoptosis in hepatoma cells

doi: 10.1038/s41598-020-64004-5

Figure Lengend Snippet: α-LA induces ER stress in hepatoma cells. ( A ) Western analysis of key players in UPR after treatment of HepG2 with 500 µM α-LA from 6 up to 48 hours. Albumin was used as loading control. For densitometric analysis protein expression was normalized to Albumin expression and values (reported over each band) have been expressed as fold change respect to control. Each lane represents a pool of three individual samples. ( B ) Schematic representation of α-LA-mediated apoptosis in hepatoma cells. Western blot images ( A ) have been cropped for clarity with full blot presented in Supplementary Fig. .

Article Snippet: The rat hepatoma cell line, FaO, and the hepatocarcinoma cell line, HepG2, were supplied by Interlab Cell Line Collection (Servizio Biotecnologie, IST, Genova, Italy), and maintained, respectively, in Dulbecco’s medium (DMEM plus Glutamax I) (Invitrogen) and supplemented with penicillin, streptomycin and 10% heat-inactivated fetal calf-serum (FCS) (Invitrogen) in a humidified atmosphere of 5% CO 2 /95% air, at 37 °C. α-Lipoic Acid (α-LA) and Thapsigargin (TG) were purchased from Sigma (Sigma-Aldrich, Milano, Italy). α-LA, dissolved in sodium hydroxide NaOH 1 N and neutralized in medium, and TG dissolved in DMSO, were added to the culture media to the final concentrations specified in the text.

Techniques: Western Blot, Control, Expressing