Journal: Molecules

Article Title: Cytotoxic Activity of Sicilian Red- and White-Grape Seed Oils on Human Liver and Colorectal Cancer Cells

doi: 10.3390/molecules31101567

Figure Lengend Snippet: Dilution-response effect of the oils on the viability of cells after 24 h of exposure. ( A ) HepG2 cells treated with WGSO (black circles) and RGSO (red circles); ( B ) CaCo-2 cells treated with WGSO; ( C ) CaCo-2 cells treated with RGSO; ( D ) diff-CaCo-2 cells treated with WGSO (black circles) and RGSO (red circles). The error bars correspond to the standard error of the mean (s.e.m.) of biological triplicates. One-way ANOVA followed by Holm–Sidak post hoc test was used. * p < 0.05; ** p < 0.001 compared to control. Normality test passed.

Article Snippet: HepG2 liver tumor cells (RRID: CVCL_0027) and CaCo-2 colorectal cancer cells (RRID: CVCL_0025), primarily obtained from ATCC (Manassas, VA, USA) and sourced from the laboratory’s internal stocks, were routinely cultured in a high-glucose Dulbecco’s Modified Eagle’s medium (DMEM; D6429, Sigma, St. Louis, MO, USA) enriched with 10% heat-inactivated fetal bovine serum (FBS, F4135, Sigma) and antibiotics (100 U/mL penicillin and 100 mg/mL streptomycin; Capricorn Scientific GmbH, Ebsdorfergrund, Germany) within a humidified environment at 37 °C under a 5% CO 2 atmosphere.

Techniques: Control

Journal: Molecules

Article Title: Cytotoxic Activity of Sicilian Red- and White-Grape Seed Oils on Human Liver and Colorectal Cancer Cells

doi: 10.3390/molecules31101567

Figure Lengend Snippet: Representative density plots for FSC-A and SSC-A in HepG2 cells ( A , B ) and CaCo-2 cells ( C – F ) cultured in control conditions ( A , C , E ) or exposed to the ID 50 of WGSO ( B , D ) or RGSO ( F ) for 24 h. ( G ) Bar graph showing the percentage of FSC-A and SSC-A values compared to control conditions normalized to 1. Values are expressed as mean % ± standard error of the mean (s.e.m). Each bar is representative of biological triplicates. One-way ANOVA followed by Holm–Sidak comparison procedure was used. Normality test passed. * p < 0.05, ** p < 0.001 compared to control.

Article Snippet: HepG2 liver tumor cells (RRID: CVCL_0027) and CaCo-2 colorectal cancer cells (RRID: CVCL_0025), primarily obtained from ATCC (Manassas, VA, USA) and sourced from the laboratory’s internal stocks, were routinely cultured in a high-glucose Dulbecco’s Modified Eagle’s medium (DMEM; D6429, Sigma, St. Louis, MO, USA) enriched with 10% heat-inactivated fetal bovine serum (FBS, F4135, Sigma) and antibiotics (100 U/mL penicillin and 100 mg/mL streptomycin; Capricorn Scientific GmbH, Ebsdorfergrund, Germany) within a humidified environment at 37 °C under a 5% CO 2 atmosphere.

Techniques: Cell Culture, Control, Comparison

Journal: Molecules

Article Title: Cytotoxic Activity of Sicilian Red- and White-Grape Seed Oils on Human Liver and Colorectal Cancer Cells

doi: 10.3390/molecules31101567

Figure Lengend Snippet: Effect of WGSO and RGSO ID 50 administration for 24 h on HepG2 and CaCo-2 cell cycle status. ( A ) Bar graph showing the percentage of cells in the subG 0 /G 1 fraction in control and treated conditions. Values are expressed as mean % ± standard error of the mean (s.e.m). ( B ) Bar graph showing the treatment-induced changes in the cell cycle phase distribution for the surviving cell population. Values are expressed as mean fold change ± standard error of the mean (s.e.m.) compared to the controls. Each bar is representative of biological triplicates. One-way ANOVA followed by Holm–Sidak comparison procedure was used. Normality test passed. * p < 0.05, ** p < 0.001 compared to control.

Article Snippet: HepG2 liver tumor cells (RRID: CVCL_0027) and CaCo-2 colorectal cancer cells (RRID: CVCL_0025), primarily obtained from ATCC (Manassas, VA, USA) and sourced from the laboratory’s internal stocks, were routinely cultured in a high-glucose Dulbecco’s Modified Eagle’s medium (DMEM; D6429, Sigma, St. Louis, MO, USA) enriched with 10% heat-inactivated fetal bovine serum (FBS, F4135, Sigma) and antibiotics (100 U/mL penicillin and 100 mg/mL streptomycin; Capricorn Scientific GmbH, Ebsdorfergrund, Germany) within a humidified environment at 37 °C under a 5% CO 2 atmosphere.

Techniques: Control, Comparison

Journal: Molecules

Article Title: Cytotoxic Activity of Sicilian Red- and White-Grape Seed Oils on Human Liver and Colorectal Cancer Cells

doi: 10.3390/molecules31101567

Figure Lengend Snippet: Effect of WGSO or RGSO ID 50 administration for 4, 16 and 24 h on the Bcl-2/Bax protein ratio in HepG2 cells ( A ) and CaCo-2 cells ( B ). The original immunoblots are shown in . Each bar is representative of technical triplicates. Values are expressed as mean fold change ± standard error of the mean (s.e.m.) compared to the controls. One-way ANOVA followed by Holm–Sidak comparison procedure was used. Normality test passed. * p < 0.001 compared to control.

Article Snippet: HepG2 liver tumor cells (RRID: CVCL_0027) and CaCo-2 colorectal cancer cells (RRID: CVCL_0025), primarily obtained from ATCC (Manassas, VA, USA) and sourced from the laboratory’s internal stocks, were routinely cultured in a high-glucose Dulbecco’s Modified Eagle’s medium (DMEM; D6429, Sigma, St. Louis, MO, USA) enriched with 10% heat-inactivated fetal bovine serum (FBS, F4135, Sigma) and antibiotics (100 U/mL penicillin and 100 mg/mL streptomycin; Capricorn Scientific GmbH, Ebsdorfergrund, Germany) within a humidified environment at 37 °C under a 5% CO 2 atmosphere.

Techniques: Western Blot, Comparison, Control

Journal: Molecules

Article Title: Cytotoxic Activity of Sicilian Red- and White-Grape Seed Oils on Human Liver and Colorectal Cancer Cells

doi: 10.3390/molecules31101567

Figure Lengend Snippet: Flow cytometric assays for externalization of phosphatidylserine in HepG2 and CaCo-2 cells cultured in control conditions or exposed to the ID 50 of the oils for 24 h. The plots show the results of representative experiments and the percentages, indicated as the mean ± standard error of the mean (s.e.m.) of biological triplicates, refer to viable annexinV-negative/PI-negative cells (bottom-left quadrant), early-apoptotic annexinV-positive/PI-negative cells (bottom-right quadrant), late-apoptotic annexinV-positive/PI-positive cells (top-right quadrant), and necrotic annexinV-negative/PI-positive cells (top-left quadrant).

Article Snippet: HepG2 liver tumor cells (RRID: CVCL_0027) and CaCo-2 colorectal cancer cells (RRID: CVCL_0025), primarily obtained from ATCC (Manassas, VA, USA) and sourced from the laboratory’s internal stocks, were routinely cultured in a high-glucose Dulbecco’s Modified Eagle’s medium (DMEM; D6429, Sigma, St. Louis, MO, USA) enriched with 10% heat-inactivated fetal bovine serum (FBS, F4135, Sigma) and antibiotics (100 U/mL penicillin and 100 mg/mL streptomycin; Capricorn Scientific GmbH, Ebsdorfergrund, Germany) within a humidified environment at 37 °C under a 5% CO 2 atmosphere.

Techniques: Cell Culture, Control

Journal: Molecules

Article Title: Cytotoxic Activity of Sicilian Red- and White-Grape Seed Oils on Human Liver and Colorectal Cancer Cells

doi: 10.3390/molecules31101567

Figure Lengend Snippet: Western blot analysis for the activation of caspase-3 in control and oil-treated HepG2 and CaCo-2 cells. The complete immunoblots are shown in .

Article Snippet: HepG2 liver tumor cells (RRID: CVCL_0027) and CaCo-2 colorectal cancer cells (RRID: CVCL_0025), primarily obtained from ATCC (Manassas, VA, USA) and sourced from the laboratory’s internal stocks, were routinely cultured in a high-glucose Dulbecco’s Modified Eagle’s medium (DMEM; D6429, Sigma, St. Louis, MO, USA) enriched with 10% heat-inactivated fetal bovine serum (FBS, F4135, Sigma) and antibiotics (100 U/mL penicillin and 100 mg/mL streptomycin; Capricorn Scientific GmbH, Ebsdorfergrund, Germany) within a humidified environment at 37 °C under a 5% CO 2 atmosphere.

Techniques: Western Blot, Activation Assay, Control

Journal: Molecules

Article Title: Cytotoxic Activity of Sicilian Red- and White-Grape Seed Oils on Human Liver and Colorectal Cancer Cells

doi: 10.3390/molecules31101567

Figure Lengend Snippet: Effect of WGSO or RGSO ID 50 administration for 4, 16 and 24 h on the cleaved caspase-1/full-length caspase-1 ratio ( A , B ) and the cleaved gasdermin-D/gasdermin-D ratio ( C , D ) in HepG2 cells ( A , C ) and CaCo-2 cells ( B , D ). The original immunoblots are shown in . Each bar is representative of technical triplicates. Values are expressed as mean fold change ± standard error of the mean (s.e.m.) compared to the controls. One-way ANOVA followed by Holm–Sidak comparison procedure was used. Normality test passed. * p < 0.001 compared to control.

Article Snippet: HepG2 liver tumor cells (RRID: CVCL_0027) and CaCo-2 colorectal cancer cells (RRID: CVCL_0025), primarily obtained from ATCC (Manassas, VA, USA) and sourced from the laboratory’s internal stocks, were routinely cultured in a high-glucose Dulbecco’s Modified Eagle’s medium (DMEM; D6429, Sigma, St. Louis, MO, USA) enriched with 10% heat-inactivated fetal bovine serum (FBS, F4135, Sigma) and antibiotics (100 U/mL penicillin and 100 mg/mL streptomycin; Capricorn Scientific GmbH, Ebsdorfergrund, Germany) within a humidified environment at 37 °C under a 5% CO 2 atmosphere.

Techniques: Western Blot, Comparison, Control

Journal: Molecules

Article Title: Cytotoxic Activity of Sicilian Red- and White-Grape Seed Oils on Human Liver and Colorectal Cancer Cells

doi: 10.3390/molecules31101567

Figure Lengend Snippet: Effect of WGSO or RGSO ID 50 administration for 4, 16 and 24 h on pMLKL protein levels in HepG2 ( A ) and CaCo-2 cells ( B ), determined by Western blot. The complete immunoblots are shown in . Each bar is representative of technical triplicates. Values are expressed as mean fold change ± standard error of the mean (s.e.m.) compared to the controls. One-way ANOVA followed by Holm–Sidak comparison procedure was used. Normality test passed. * p < 0.05, ** p < 0.001 compared to control.

Article Snippet: HepG2 liver tumor cells (RRID: CVCL_0027) and CaCo-2 colorectal cancer cells (RRID: CVCL_0025), primarily obtained from ATCC (Manassas, VA, USA) and sourced from the laboratory’s internal stocks, were routinely cultured in a high-glucose Dulbecco’s Modified Eagle’s medium (DMEM; D6429, Sigma, St. Louis, MO, USA) enriched with 10% heat-inactivated fetal bovine serum (FBS, F4135, Sigma) and antibiotics (100 U/mL penicillin and 100 mg/mL streptomycin; Capricorn Scientific GmbH, Ebsdorfergrund, Germany) within a humidified environment at 37 °C under a 5% CO 2 atmosphere.

Techniques: Western Blot, Comparison, Control

Journal: Molecules

Article Title: Cytotoxic Activity of Sicilian Red- and White-Grape Seed Oils on Human Liver and Colorectal Cancer Cells

doi: 10.3390/molecules31101567

Figure Lengend Snippet: Effect of WGSO or RGSO ID 50 on the amount of HMGB1 protein released in the media of HepG2 and CaCo-2 cells after 24 h, determined by dot blot. The original immunoblot is shown in . Each bar is representative of technical triplicates. Values are expressed as mean fold change ± standard error of the mean (s.e.m.) compared to the controls. One-way ANOVA followed by Holm–Sidak comparison procedure was used. Normality test passed. * p < 0.001 compared to control.

Article Snippet: HepG2 liver tumor cells (RRID: CVCL_0027) and CaCo-2 colorectal cancer cells (RRID: CVCL_0025), primarily obtained from ATCC (Manassas, VA, USA) and sourced from the laboratory’s internal stocks, were routinely cultured in a high-glucose Dulbecco’s Modified Eagle’s medium (DMEM; D6429, Sigma, St. Louis, MO, USA) enriched with 10% heat-inactivated fetal bovine serum (FBS, F4135, Sigma) and antibiotics (100 U/mL penicillin and 100 mg/mL streptomycin; Capricorn Scientific GmbH, Ebsdorfergrund, Germany) within a humidified environment at 37 °C under a 5% CO 2 atmosphere.

Techniques: Dot Blot, Western Blot, Comparison, Control

Journal: Molecules

Article Title: Cytotoxic Activity of Sicilian Red- and White-Grape Seed Oils on Human Liver and Colorectal Cancer Cells

doi: 10.3390/molecules31101567

Figure Lengend Snippet: Effect of WGSO or RGSO ID 50 administration for 4, 16 and 24 h on hsp60 ( A , B ) and hsp90 ( C , D ) protein levels in HepG2 ( A , C ) and CaCo-2 cells ( B , D ), determined by Western blot. The complete immunoblots are shown in . Each bar is representative of technical triplicates. Values are expressed as mean fold change ± standard error of the mean (s.e.m.) compared to the controls. One-way ANOVA followed by Holm–Sidak comparison procedure was used. Normality test passed. * p < 0.001 compared to control.

Article Snippet: HepG2 liver tumor cells (RRID: CVCL_0027) and CaCo-2 colorectal cancer cells (RRID: CVCL_0025), primarily obtained from ATCC (Manassas, VA, USA) and sourced from the laboratory’s internal stocks, were routinely cultured in a high-glucose Dulbecco’s Modified Eagle’s medium (DMEM; D6429, Sigma, St. Louis, MO, USA) enriched with 10% heat-inactivated fetal bovine serum (FBS, F4135, Sigma) and antibiotics (100 U/mL penicillin and 100 mg/mL streptomycin; Capricorn Scientific GmbH, Ebsdorfergrund, Germany) within a humidified environment at 37 °C under a 5% CO 2 atmosphere.

Techniques: Western Blot, Comparison, Control

Journal: Molecules

Article Title: Cytotoxic Activity of Sicilian Red- and White-Grape Seed Oils on Human Liver and Colorectal Cancer Cells

doi: 10.3390/molecules31101567

Figure Lengend Snippet: Representative flow cytometric profiles for ROS production in HepG2 cells ( A , B ) and CaCo-2 cells ( C , D ) cultured in control conditions or exposed to the ID 50 of WGSO or RGSO for 4 ( A , C ) or 16 h ( B , D ). ( E ) Bar graph showing the MFI of ROS indicator, obtained from triplicate experiments. Cumulative data from flow cytometry experiments were analyzed for the MFI of each condition; data normalization was then achieved by dividing the MFI of the treated cell samples by the MFI of the reference controls. Each bar is representative of biological triplicates. Values are expressed as mean fold change ± standard error of the mean (s.e.m.) compared to the controls. One-way ANOVA followed by Holm–Sidak comparison procedure was used. Normality test passed. * p < 0.01, ** p < 0.05, *** p < 0.001 compared to control.

Article Snippet: HepG2 liver tumor cells (RRID: CVCL_0027) and CaCo-2 colorectal cancer cells (RRID: CVCL_0025), primarily obtained from ATCC (Manassas, VA, USA) and sourced from the laboratory’s internal stocks, were routinely cultured in a high-glucose Dulbecco’s Modified Eagle’s medium (DMEM; D6429, Sigma, St. Louis, MO, USA) enriched with 10% heat-inactivated fetal bovine serum (FBS, F4135, Sigma) and antibiotics (100 U/mL penicillin and 100 mg/mL streptomycin; Capricorn Scientific GmbH, Ebsdorfergrund, Germany) within a humidified environment at 37 °C under a 5% CO 2 atmosphere.

Techniques: Cell Culture, Control, Flow Cytometry, Comparison

Journal: Molecules

Article Title: Cytotoxic Activity of Sicilian Red- and White-Grape Seed Oils on Human Liver and Colorectal Cancer Cells

doi: 10.3390/molecules31101567

Figure Lengend Snippet: Representative flow cytometric profiles for AVOs’ accumulation in HepG2 cells ( A , B ) and CaCo-2 cells ( C , D ) cultured in control conditions or exposed to the ID 50 of WGSO or RGSO for 4 ( A , C ) or 16 h ( B , D ). ( E ) Bar graph showing the mean fluorescence index (MFI) of acridine orange stain, obtained from triplicate experiments. Cumulative data from flow cytometry experiments were analyzed for the MFI of each condition; data normalization was then achieved by dividing the MFI of the treated cell samples by the MFI of the reference controls. Each bar is representative of biological triplicates. Values are expressed as mean fold change ± standard error of the mean (s.e.m.) compared to the controls. One-way ANOVA followed by Holm–Sidak comparison procedure was used. Normality test passed. * p < 0.001 compared to control.

Article Snippet: HepG2 liver tumor cells (RRID: CVCL_0027) and CaCo-2 colorectal cancer cells (RRID: CVCL_0025), primarily obtained from ATCC (Manassas, VA, USA) and sourced from the laboratory’s internal stocks, were routinely cultured in a high-glucose Dulbecco’s Modified Eagle’s medium (DMEM; D6429, Sigma, St. Louis, MO, USA) enriched with 10% heat-inactivated fetal bovine serum (FBS, F4135, Sigma) and antibiotics (100 U/mL penicillin and 100 mg/mL streptomycin; Capricorn Scientific GmbH, Ebsdorfergrund, Germany) within a humidified environment at 37 °C under a 5% CO 2 atmosphere.

Techniques: Cell Culture, Control, Fluorescence, Staining, Flow Cytometry, Comparison

Journal: Molecules

Article Title: Cytotoxic Activity of Sicilian Red- and White-Grape Seed Oils on Human Liver and Colorectal Cancer Cells

doi: 10.3390/molecules31101567

Figure Lengend Snippet: Effect of WGSO ID 50 administration for 4 and 16 h on Beclin-1 ( A ) and p62 ( B ) protein levels and LC3-II/LC3-I protein ratio ( C ) in HepG2 cells, determined by Western blot. The complete immunoblots are shown in . Each bar is representative of technical triplicates. Values are expressed as mean fold change ± standard error of the mean (s.e.m.) compared to the controls. One-way ANOVA followed by Holm–Sidak comparison procedure was used. Normality test passed. * p < 0.05, ** p < 0.001 compared to control.

Article Snippet: HepG2 liver tumor cells (RRID: CVCL_0027) and CaCo-2 colorectal cancer cells (RRID: CVCL_0025), primarily obtained from ATCC (Manassas, VA, USA) and sourced from the laboratory’s internal stocks, were routinely cultured in a high-glucose Dulbecco’s Modified Eagle’s medium (DMEM; D6429, Sigma, St. Louis, MO, USA) enriched with 10% heat-inactivated fetal bovine serum (FBS, F4135, Sigma) and antibiotics (100 U/mL penicillin and 100 mg/mL streptomycin; Capricorn Scientific GmbH, Ebsdorfergrund, Germany) within a humidified environment at 37 °C under a 5% CO 2 atmosphere.

Techniques: Western Blot, Comparison, Control

Journal: Frontiers in Oncology

Article Title: A novel auto-fluorescent porphyrin-lipid nanoparticle strategy for CTNNB1 gene silencing in hepatocellular carcinoma

doi: 10.3389/fonc.2026.1779803

Figure Lengend Snippet: Porphyrin-LNP synthesis and characterization. (A) Porphyrin-LNPs were synthesized via microfluidic rapid mixing. (B) . Cryo-TEM image of porphyrin-LNP (scale bar = 100 nm). (C) Size distribution of porphyrin-LNP-si-Scramble and (D) porphyrin-LNP-si-CTNNB1 measured by dynamic light scattering (DLS). (E) Confocal imaging of FAM-siRNA delivery by porphyrin-LNP. Hep3B and HepG2 cells were treated with porphyrin-LNP encapsulating FAM-siRNA (200 nM, ~2 μM porphyrin) for 6 h. Confocal images show FAM-siRNA (green) and porphyrin (magenta).

Article Snippet: The Hep3B (HB-8064) and HepG2 (HB-8065) human liver cancer cell lines were obtained from the American Type Culture Collection (ATCC) and tested negative for mycoplasma contamination.

Techniques: Synthesized, Imaging

Journal: Frontiers in Oncology

Article Title: A novel auto-fluorescent porphyrin-lipid nanoparticle strategy for CTNNB1 gene silencing in hepatocellular carcinoma

doi: 10.3389/fonc.2026.1779803

Figure Lengend Snippet: Validation of CTNNB1 knockdown efficiency using porphyrin-LNP-siRNA. Downregulation of CTNNB1 expression after transfection of porphyrin-LNP-siRNA targeting CTNNB1 in (A) Hep3B and (B) HepG2 cells. **** p < 0.0001; one-way ANOVA test. Reduced colony formation after transfection of porphyrin-LNP-siRNA targeting CTNNB1 in (C) Hep3B and (D) HepG2 cells. * p < 0.05, *** p < 0.001, one-way ANOVA test.

Article Snippet: The Hep3B (HB-8064) and HepG2 (HB-8065) human liver cancer cell lines were obtained from the American Type Culture Collection (ATCC) and tested negative for mycoplasma contamination.

Techniques: Biomarker Discovery, Knockdown, Expressing, Transfection

Journal: bioRxiv

Article Title: Methionine Deprivation-induced Reprogramming of Hepatic Rhythms Is Mediated by Glucocorticoid Receptor

doi: 10.64898/2026.05.04.722637

Figure Lengend Snippet: A. Immunoblot of global H3 methylation levels in livers from mice fed on control and MD diet for three weeks. B. Immunoblot of H3 methylation levels in HepG2 cultured in medium with indicated methionine concentration for 24 hr. C. The Venn diagram depicts rhythmic H3K36me3 peaks in mice liver on control diet and MD diet measured by CUT&Tag. Rhythmic peaks were defined by comparing enrichment signals between ZT9 and ZT21 as described in Methods. D. Metagene profiles depicting H3K36me3 enrichment for 453 peaks with a stronger signal at ZT21 compared to ZT9 in control-diet fed mice (peak center ± 10 kb). E. IGV browser tracks showing H3K36me3 enrichment at Elovl3 and Elovl5 genes. Diet and Zeitgeber time for animal collection are indicated to the left, gene model is indicated below the data. F. TMM normalized expression values are presented in the graph for the Elovl5 gene. G. TMM normalized expression values are presented in the graph for the Elovl3 gene. H. The Venn diagram depicts rhythmic H3K4me3 peaks in mice liver on control diet and MD diet. Rhythmic peaks were defined by comparing enrichment signals between ZT9 and ZT21 as described in Methods. I. IGV browser tracks showing H3K4me3 and H3K36me3 enrichment at Insig2 gene. Diet and Zeitgeber time for animal collection are indicated to the left, gene model is indicated below the data. J. TMM normalized expression values are presented in the graph for the Insig2 gene. Diet group and Zeitgeber time are indicated on the graphs. See also Figure S4

Article Snippet: Human liver cancer cell HepG2 was obtained from ATCC and maintained in RPMI-1640 (GIBCO, 11875119) supplemented with 10% FBS at 37 °C with 5% CO 2 .

Techniques: Western Blot, Methylation, Control, Cell Culture, Concentration Assay, Expressing

Journal: bioRxiv

Article Title: Methionine Deprivation-induced Reprogramming of Hepatic Rhythms Is Mediated by Glucocorticoid Receptor

doi: 10.64898/2026.05.04.722637

Figure Lengend Snippet: A. Serum levels of corticosterone of six-month old male mice after three weeks on the indicated diet. The column graphs indicate mean value for 3-5 animals at indicated zeitgeber time points. Significance (**p<0.01, t test) is indicated with asterisks. B. The table shows the number of oscillating GR in liver from mice on indicated diet for three weeks. C. The Venn diagram depicts the rhythmic GR binding peaks in mice liver on control diet and MD diet detected by CUT&RUN. The rhythmic peaks were defined by comparing the enrichment signals between ZT9 and ZT21 as described in Methods. D. IGV browser tracks showing the GR enrichment at Insig2 gene. Diet and Zeitgeber time for animal collection are indicated to the left. E. Motif analysis for 3,967 GR binding peaks with a stronger signal at ZT9 compared to ZT21 in control-diet fed mice. F. Motif analysis for 2,556 GR binding peaks with a stronger signal at ZT9 compared to ZT21 in MD diet fed mice. G. On the left, the western blot depicts the protein levels of STAT5 and GR in whole cell lysis of liver. GAPDH is included as the loading control. On the right, the western blot depicts the phosphorylation levels of STAT5, protein levels of STAT5 and GR in the nuclei extraction from liver. Lamin A/C is included as the loading control. H. TMM normalized expression values are presented in the graph for the Egfr gene. Diet group and Zeitgeber time are indicated on the graphs. I. The western blot depicts the protein levels of EGFR in liver from mice after three weeks on the indicated diet. GAPDH is included as the loading control. J. The western blot depicts the protein levels of EGFR and STAT5 in whole cell lysis and nuclei extraction of HepG2 cultured in medium with high methionine concentration (100 µM) or low methionine concentration (10 µM) for 40 hours following treated with EGF (0, 100 and 200 ng/mL) for 30 min. GAPDH and Lamin A/C are included as loading control. See also Figure S5

Article Snippet: Human liver cancer cell HepG2 was obtained from ATCC and maintained in RPMI-1640 (GIBCO, 11875119) supplemented with 10% FBS at 37 °C with 5% CO 2 .

Techniques: Binding Assay, Control, Western Blot, Lysis, Phospho-proteomics, Extraction, Expressing, Cell Culture, Concentration Assay

Journal: Vaccines

Article Title: Establishment and Validation of a Cell-Based Relative Potency Method for Respiratory Syncytial Virus mRNA Vaccine Drug Substance

doi: 10.3390/vaccines14050401

Figure Lengend Snippet: Optimization of cell-related conditions for the RSV mRNA potency assay. ( A ) Dose–response curves of RSV mRNA transfection in different cell lines (293T, Vero, HEp-2, HepG2, and A549). Cells were transfected with increasing amounts of RSV mRNA (0.0002–0.48 µg/well), and RSV prefusion F protein expression was measured by ELISA as OD 450 –OD 630 . The corresponding EC 50 and slope values are summarized in the inset table. ( B ) Effect of HepG2 cell seeding density on RSV prefusion F protein expression. HepG2 cells were seeded at 0.5 × 10 5 , 1.0 × 10 5 , or 2.0 × 10 5 cells/well and transfected with increasing amounts of RSV mRNA (0.0002–0.48 µg/well). Protein expression was measured by ELISA as OD 450 –OD 630 .

Article Snippet: Cell lines used for mRNA transfection included the human liver carcinoma cell line HepG2 (American Type Culture Collection, ATCC, Manassas, VA, USA), human non-small cell lung carcinoma cell line A549 (ATCC), African green monkey kidney epithelial cell line Vero (ATCC), human epidermoid carcinoma type 2 cell line Hep-2 (ATCC), and human embryonic kidney cell line 293T (China Center for Type Culture Collection, CCTCC, Wuhan, China).

Techniques: Potency Assay, Transfection, Expressing, Enzyme-linked Immunosorbent Assay