Journal: Experimental & Molecular Medicine

Article Title: Antitumorigenic and antiangiogenic efficacy of apatinib in liver cancer evaluated by multimodality molecular imaging

doi: 10.1038/s12276-019-0274-7

Figure Lengend Snippet: Half-maximal inhibitory concentration (IC 50 ) of apatinib and sorafenib in hepatocellular carcinoma cell lines

Article Snippet: The HepG2-Red-fLuc cell line was purchased from PerkinElmer (Waltham, MA, USA).

Techniques: Concentration Assay

Journal: Experimental & Molecular Medicine

Article Title: Antitumorigenic and antiangiogenic efficacy of apatinib in liver cancer evaluated by multimodality molecular imaging

doi: 10.1038/s12276-019-0274-7

Figure Lengend Snippet: a HepG2-Red-fLuc and ( b ) SMMC-7721-fLuc cell-derived tumor-bearing mice on days 0, 6, 12, and 18 post-drug treatment. Changes in BLI signal intensity, tumor volume, and mouse body weight in mice bearing HepG2-Red-fLuc ( c, e, g ) and SMMC-7721-fLuc ( d, f, h ) cell xenograft tumors were evaluated. (*) P < 0.05, (**) P < 0.01, (***) P < 0.001 vs. the control group

Article Snippet: The HepG2-Red-fLuc cell line was purchased from PerkinElmer (Waltham, MA, USA).

Techniques: Derivative Assay, Control

Journal: Experimental & Molecular Medicine

Article Title: Antitumorigenic and antiangiogenic efficacy of apatinib in liver cancer evaluated by multimodality molecular imaging

doi: 10.1038/s12276-019-0274-7

Figure Lengend Snippet: a BLI signals of orthotopic HepG2-Red-fLuc cell-derived tumor-bearing mice on days 0, 3, 6, 9, 12, 15, and 18 post-drug treatment. b BLI signal intensity of mice. (*) P < 0.05, (**) P < 0.01 vs. the control group. c Body weight of mice. (*) P < 0.05, day 6 vs. day 0 in the sorafenib group. d The 3D BLT reconstruction in the control and sorafenib and apatinib treatment groups on day 20 post-drug treatment

Article Snippet: The HepG2-Red-fLuc cell line was purchased from PerkinElmer (Waltham, MA, USA).

Techniques: Derivative Assay, Control

Journal: Antioxidants

Article Title: Cell-Based Antioxidant Properties and Synergistic Effects of Natural Plant and Algal Extracts Pre and Post Intestinal Barrier Transport

doi: 10.3390/antiox11030565

Figure Lengend Snippet: Comparison of AOP1 EC 50 s evaluated in Caco-2 and HepG2 cells for the 12 natural extracts selected for the intestinal barrier transport study. Extracts were selected according to the best EC 50 s obtained with AOP1 assay. In order to increase the diversity of tested extracts, some Vitis vinifera extracts were removed from the list because of redundancy with each other. ND = not determined (no full effect observed).

Article Snippet: The HepG2/Nrf2 cells were treated with conditions for 17 h; then, they were treated with a mix (BPS Bioscience, USA) comprising cell lysis solution and luciferin (substrate of luciferase) for 40 min. Luminescence was read on a Varioskan Flash Spectral Scanning Multimode Reader.

Techniques:

Journal: Antioxidants

Article Title: Cell-Based Antioxidant Properties and Synergistic Effects of Natural Plant and Algal Extracts Pre and Post Intestinal Barrier Transport

doi: 10.3390/antiox11030565

Figure Lengend Snippet: Indirect antioxidant ARE/Nrf2 assay data obtained for Syzygium aromaticum extract (given as an example) in HepG2/Nrf2 cells. All other extracts were subjected to the same protocol and data analyses. ( A ) Assay of increasing concentrations showing the typical bell-shaped profile (see text for explanation). Sulforaphane 25 µM was used as positive control; ( B ) dose–response and sigmoid fit curve used for Efficacy Concentration (EC) estimations.

Article Snippet: The HepG2/Nrf2 cells were treated with conditions for 17 h; then, they were treated with a mix (BPS Bioscience, USA) comprising cell lysis solution and luciferin (substrate of luciferase) for 40 min. Luminescence was read on a Varioskan Flash Spectral Scanning Multimode Reader.

Techniques: Positive Control, Concentration Assay

Journal: Antioxidants

Article Title: Cell-Based Antioxidant Properties and Synergistic Effects of Natural Plant and Algal Extracts Pre and Post Intestinal Barrier Transport

doi: 10.3390/antiox11030565

Figure Lengend Snippet: ARE/Nrf2 EC 50 s established in HepG2 cells for the 12 natural extracts selected for the Caco-2 intestinal barrier transport study. ND = not determined; R 2 = determination coefficient.

Article Snippet: The HepG2/Nrf2 cells were treated with conditions for 17 h; then, they were treated with a mix (BPS Bioscience, USA) comprising cell lysis solution and luciferin (substrate of luciferase) for 40 min. Luminescence was read on a Varioskan Flash Spectral Scanning Multimode Reader.

Techniques:

Journal: Antioxidants

Article Title: Cell-Based Antioxidant Properties and Synergistic Effects of Natural Plant and Algal Extracts Pre and Post Intestinal Barrier Transport

doi: 10.3390/antiox11030565

Figure Lengend Snippet: Effect of post-intestinal barrier transport compartments on ARE/NRF2 pathway activation in HepG2 cells. Results are expressed as fold increase compared to constitutive medium ARE/Nrf2 gene expression. Values represent means of quadruplicates obtained from two independent intestinal barrier transport experiments. Sulforaphane 25 µM data is provided as positive control; *** p < 0.005.

Article Snippet: The HepG2/Nrf2 cells were treated with conditions for 17 h; then, they were treated with a mix (BPS Bioscience, USA) comprising cell lysis solution and luciferin (substrate of luciferase) for 40 min. Luminescence was read on a Varioskan Flash Spectral Scanning Multimode Reader.

Techniques: Activation Assay, Expressing, Positive Control

Journal: Antioxidants

Article Title: Cell-Based Antioxidant Properties and Synergistic Effects of Natural Plant and Algal Extracts Pre and Post Intestinal Barrier Transport

doi: 10.3390/antiox11030565

Figure Lengend Snippet: Effect of combinations of post-intestinal barrier compartments on ARE/NRF2 pathway in HepG2 cells. Results are expressed as ARE/Nrf2 gene expression fold increase compared to constitutive medium. Values represent means of pentaplicates for all 50/50 combinations and of triplicates for all others. All data were normalized to constitutive medium values; *** p < 0.001; ** p < 0.01.

Article Snippet: The HepG2/Nrf2 cells were treated with conditions for 17 h; then, they were treated with a mix (BPS Bioscience, USA) comprising cell lysis solution and luciferin (substrate of luciferase) for 40 min. Luminescence was read on a Varioskan Flash Spectral Scanning Multimode Reader.

Techniques: Expressing

Journal: Antioxidants

Article Title: Cell-Based Antioxidant Properties and Synergistic Effects of Natural Plant and Algal Extracts Pre and Post Intestinal Barrier Transport

doi: 10.3390/antiox11030565

Figure Lengend Snippet: Synergistic effects on ARE/Nrf2 gene expression of post-intestinal barrier compartment combinations. Combin. FI = gene expression fold increase (FI) measure for the combination; S indep. FI = sum of the increase in gene expression fold increase (FI) measures of each of the two extracts taken independently as depicted in Figure 6 ; Var. (%) = variation in percentage of the two values. A positive percentage value indicates a synergistic effect.

Article Snippet: The HepG2/Nrf2 cells were treated with conditions for 17 h; then, they were treated with a mix (BPS Bioscience, USA) comprising cell lysis solution and luciferin (substrate of luciferase) for 40 min. Luminescence was read on a Varioskan Flash Spectral Scanning Multimode Reader.

Techniques: 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: bioRxiv

Article Title: Sterols from cholesterol synthesis control distinct gene regulatory pathways

doi: 10.1101/2023.05.19.538399

Figure Lengend Snippet: Proliferation and cell cycle analysis of HepG2 cell lines. Plots represent the proliferation of HepG2 cell lines in different serum and cholesterol conditions on A) FBS , B) LDS + 30 µg/ml cholesterol and C) LDS . The assay was performed using the CCK8 method and data are represented as mean +/- SE (n=6). Statistical significance was tested using One-way ANOVA with comparison to control, the Native cell line result at the same time point, p-value - *< 0.05, **< 0.01, ***< 0.001, ****<0.0001. Cell cycle measurements using propidium iodide are represented on D) as a direct measurement and on E) results normalized to total cell number (100%) and statistically evaluated. All KOs were compared with control Native cell line using the statistical test described above.

Article Snippet: HepG2 (human liver cancer cell line, Synthego, Menalo Park, CA, USA) with CRISPR-Cas9 mediated deletion of one of the enzymes from cholesterol synthesis (CYP51A1, DHCR24 and SC5D) and the Native cell line were ordered from Synthego (Synthego, Menalo Park, CA, USA).

Techniques: Cell Cycle Assay

Journal: bioRxiv

Article Title: Sterols from cholesterol synthesis control distinct gene regulatory pathways

doi: 10.1101/2023.05.19.538399

Figure Lengend Snippet: Expression of mRNA and proteins in control HepG2 cells (Native) and in knockout cells of enzymes from the late part of cholesterol synthesis ( CYP51 KO, DHCR24 KO, SCD KO). Bars represent the mean + SD of three measurements. A – C: Relative mRNA expression of targeted genes ( A - CYP51 , B - DHCR24 , C - SC5D ). Expression data was normalized to ACTB , GAPDH, and RPLP0 reference genes. D - I : Targeted protein expression ( D - CYP51A1, E - DHCR24, F - SC5D, G – HMGCR , H – c-SREBP2 , I – n-SREBP2), by western blotting and appropriate antibody as described in Methods. 10 µg of proteins were loaded and normalized to total proteins. J) Sterol analysis by LC-MS. Sterol concentrations are represented as a mean +/- SD in ng/10 7 cells, except for cholesterol in µg/10 7 cells. The colours represent relative concentration compared to Native cells (Blue – Depleted; Red – Accumulated), ND – non-detectable, N=3. Cholesterol* in KO cells results from the culture medium. K) A simplified cholesterol synthesis, with measured sterols from the post-lanosterol part of a synthesis and the position of deleted enzymes in red. The Bloch and Kandutsch-Russell sterol pathways are indicated. For statistics, one-way ANOVA was used. *p<0.1, **p<0.05, ***p<0.01, ****p<0.001.

Article Snippet: HepG2 (human liver cancer cell line, Synthego, Menalo Park, CA, USA) with CRISPR-Cas9 mediated deletion of one of the enzymes from cholesterol synthesis (CYP51A1, DHCR24 and SC5D) and the Native cell line were ordered from Synthego (Synthego, Menalo Park, CA, USA).

Techniques: Expressing, Knock-Out, Western Blot, Liquid Chromatography with Mass Spectroscopy, Concentration Assay

Journal: bioRxiv

Article Title: Sterols from cholesterol synthesis control distinct gene regulatory pathways

doi: 10.1101/2023.05.19.538399

Figure Lengend Snippet: Differential gene expression (DEG) using Clarium S microarrays. A) PCA (Principal component analysis) of raw gene expression data for all four conditions (Native, CYP51 KO, DHCR24 KO, and SC5D KO), each measurement in 3 biological replicates. B) Number of up, down, and total differently expressed genes in all three genotypes compare to the Native HepG2 cell line after False discovery rate (FDR) correction (adj pvalue<0.05). C) Venn diagram of DEGs comparing gene expression between different genotypes. D) Venn diagram of changed KEGG pathways comparing different genotypes. E) ( CYP51 KO), F) ( DHCR24 KO) and G) ( SC5D KO) represent volcano plots of differentially expressed genes, downregulated shown in blue and upregulated in red (FDR<0.05). The top 10 statistically significant genes are labelled. H) ( CYP51 KO), I) ( DHCR24 KO) and J) ( SC5D KO) represent changes in KEGG metabolic pathways (FDR<0.05), with red showing the pathways that are upregulated and blue those that are downregulated. The # after the pathway name indicates significant change unique for this KOs cell line.

Article Snippet: HepG2 (human liver cancer cell line, Synthego, Menalo Park, CA, USA) with CRISPR-Cas9 mediated deletion of one of the enzymes from cholesterol synthesis (CYP51A1, DHCR24 and SC5D) and the Native cell line were ordered from Synthego (Synthego, Menalo Park, CA, USA).

Techniques: Expressing

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