Journal: Frontiers in Microbiology

Article Title: Identification of Chebulinic Acid and Chebulagic Acid as Novel Influenza Viral Neuraminidase Inhibitors

doi: 10.3389/fmicb.2020.00182

Figure Lengend Snippet: Reporter luciferase-based one-cycle infection inhibition assay. MDCK cells growing in 96-well plates were infected with reporter influenza PR8-PB2-Gluc virus at an MOI of 0.1, in the absence or presence of increasing concentrations of (A) CHLA and (B) CHLI. Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 4% fetal bovine serum (FBS) were used as culture medium during infection to avoid generation of infectious progeny viruses. At 24 h post infection (p.i.), Gluc expression was determined, and viral infectivities were evaluated. Data are means ± SD from three independent experiments. * p

Article Snippet: Cells, Viruses, and Compounds Madin–Darby canine kidney (MDCK) epithelial cells were grown in Dulbecco’s modified Eagle’s medium (DMEM; Cellgro, Manassas, VA, United States) supplemented with 10% fetal bovine serum (FBS; Gibco, Carlsbad, CA, United States), 1000 units/mL of penicillin, and 100 μg/mL of streptomycin (Invitrogen, Carlsbad, CA, United States).

Techniques: Luciferase, Infection, Inhibition, Modification, Expressing

Journal: International Journal of Oncology

Article Title: 2,3,7,8-tetrachlorodibenzo-p-dioxin suppresses the growth of human colorectal cancer cells in vitro: Implication of the aryl hydrocarbon receptor signaling

doi: 10.3892/ijo.2019.4703

Figure Lengend Snippet: TCDD suppresses colony formation in RKO human colorectal cancer cells in vitro . Cells (1×10 3 cells/well) were seeded into 6-well plates and cultured in Dulbecco’s modified Eagle’s medium containing 10% fetal bovine serum, 1% penicillin/streptomycin and 1% fungizone in the presence of vehicle (1% dimethyl sulfoxide) or TCDD (1 or 10 nM) for 5 days when visible clones formed. The colonies were washed with PBS, fixed with methanol and then stained with 0.5% crystal violet. (A) Stained cells are presented as images (×10), and (B) the colonies containing > 50 cells were counted under a microscope. * P

Article Snippet: Dulbecco’s modified Eagle’s medium (DMEM; including 4.5 g/l glucose, L-glutamine and sodium pyruvate) and antibiotics (100 µ g/ml penicillin and 100 µ g/ml streptomycin; P/S) were obtained from Corning Life Sciences (Manassas, VA, USA).

Techniques: In Vitro, Cell Culture, Modification, Clone Assay, Staining, Microscopy

Journal: International Journal of Oncology

Article Title: 2,3,7,8-tetrachlorodibenzo-p-dioxin suppresses the growth of human colorectal cancer cells in vitro: Implication of the aryl hydrocarbon receptor signaling

doi: 10.3892/ijo.2019.4703

Figure Lengend Snippet: TCDD suppresses the proliferation of RKO human colorectal cancer cells in vitro . The cells (1×10 5 cells/well in 24-well plates) were cultured in Dulbecco’s modified Eagle’s medium containing 10% fetal bovine serum, 1% penicillin/streptomycin and 1% fungizone in the presence of vehicle (1% dimethyl sulfoxide) or TCDD (0.01-100 nM) for (A) 3 or (B) 7 days. After culture, the numbers of attached cells were counted. Data are presented as mean ± standard deviation obtained from 8 wells of 2 replicate plates per dataset using different dishes and cell preparations. * P

Article Snippet: Dulbecco’s modified Eagle’s medium (DMEM; including 4.5 g/l glucose, L-glutamine and sodium pyruvate) and antibiotics (100 µ g/ml penicillin and 100 µ g/ml streptomycin; P/S) were obtained from Corning Life Sciences (Manassas, VA, USA).

Techniques: In Vitro, Cell Culture, Modification, Standard Deviation

Journal: International Journal of Oncology

Article Title: 2,3,7,8-tetrachlorodibenzo-p-dioxin suppresses the growth of human colorectal cancer cells in vitro: Implication of the aryl hydrocarbon receptor signaling

doi: 10.3892/ijo.2019.4703

Figure Lengend Snippet: The TCDD-induced increase in CYP1A1 levels are suppressed in the regucalcin-overexpressing RKO human colorectal cancer cells in vitro . The wild-type RKO cells or regucalcin-overexpressing transfectants (1×10 6 cells/dish) were cultured in Dulbecco’s modified Eagle’s medium containing 10% fetal bovine serum, 1% penicillin/streptomycin and 1% fungizone in the presence or absence of vehicle (1% dimethyl sulfoxide) or TCDD (10 nM) for 3 days and then cell lysates were centrifuged. Subsequently, 40 µ g of the supernatant protein per lane were separated by SDS-PAGE and transferred to nylon membranes for western blotting using specific antibodies against the indicated proteins. Representative data from three independent experiments using different cell preparations are presented, and data are presented as mean ± standard deviation. (A) Representative film images of the TCDD effect. (B) Presented relative to β-actin of the TCDD effect. * P

Article Snippet: Dulbecco’s modified Eagle’s medium (DMEM; including 4.5 g/l glucose, L-glutamine and sodium pyruvate) and antibiotics (100 µ g/ml penicillin and 100 µ g/ml streptomycin; P/S) were obtained from Corning Life Sciences (Manassas, VA, USA).

Techniques: In Vitro, Cell Culture, Modification, SDS Page, Western Blot, Standard Deviation

Journal: International Journal of Oncology

Article Title: 2,3,7,8-tetrachlorodibenzo-p-dioxin suppresses the growth of human colorectal cancer cells in vitro: Implication of the aryl hydrocarbon receptor signaling

doi: 10.3892/ijo.2019.4703

Figure Lengend Snippet: TCDD regulates the expression of proteins associated with AHR signaling in RKO human colorectal cancer cells in vitro . The cells (1×10 6 cells/dish) were cultured in Dulbecco’s modified Eagle’s medium containing 10% fetal bovine serum, 1% penicillin/streptomycin and 1% fungizone in the presence of vehicle (1% dimethyl sulfoxide) or TCDD (10 nM) for 3 days. Cell lysates were prepared and centrifuged, and 40 µ g of the supernatant protein per lane were separated by SDS-PAGE and transferred to nylon membranes for western blotting using specific antibodies against various proteins as indicated. Data represent a typical figure of three independent experiments using different cell preparations, and also are presented as mean ± standard deviation. (A) Representative film image for cell signaling-associated proteins. (B) Relative to β-actin cell signaling-associated protein levels. (C) Representative film image of tumor suppressor proteins.(D) Relative to β-actin tumor suppressor proteins. * P

Article Snippet: Dulbecco’s modified Eagle’s medium (DMEM; including 4.5 g/l glucose, L-glutamine and sodium pyruvate) and antibiotics (100 µ g/ml penicillin and 100 µ g/ml streptomycin; P/S) were obtained from Corning Life Sciences (Manassas, VA, USA).

Techniques: Expressing, In Vitro, Cell Culture, Modification, SDS Page, Western Blot, Standard Deviation

Journal: International Journal of Oncology

Article Title: 2,3,7,8-tetrachlorodibenzo-p-dioxin suppresses the growth of human colorectal cancer cells in vitro: Implication of the aryl hydrocarbon receptor signaling

doi: 10.3892/ijo.2019.4703

Figure Lengend Snippet: AHR and CYP1A1 levels are suppressed in regucalcin-overexpressing RKO human colorectal cancer cells in vitro . The wild-type RKO cells or regucalcin-overexpressing RKO cells (1×10 6 cells/per dish) were cultured in Dulbecco’s modified Eagle’s medium containing 10% fetal bovine serum, 1% penicillin/streptomycin and 1% fungizone in the presence or absence of vehicle (1% dimethyl sulfoxide) for 3 days. After culture, resulting cell lysates were centrifuged, and 40 µ g of the supernatant protein per lane were separated by SDS-PAGE and transferred to nylon membranes for western blotting using specific antibodies as indicated. Data represent a typical figure obtained from three independent experiments using different cell preparations, and also are presented as mean ± standard deviation. (A) Representative film image for regucalcin. (B) Relative to β-actin regucalcin level. (C) Representative film image of AHR and CYP1A1. (D) Relative to β-actin AHR and CYP1A1 levels. * P

Article Snippet: Dulbecco’s modified Eagle’s medium (DMEM; including 4.5 g/l glucose, L-glutamine and sodium pyruvate) and antibiotics (100 µ g/ml penicillin and 100 µ g/ml streptomycin; P/S) were obtained from Corning Life Sciences (Manassas, VA, USA).

Techniques: In Vitro, Cell Culture, Modification, SDS Page, Western Blot, Standard Deviation

Journal: Drug Design, Development and Therapy

Article Title: (+)-Grandifloracin, an antiausterity agent, induces autophagic PANC-1 pancreatic cancer cell death

doi: 10.2147/DDDT.S52168

Figure Lengend Snippet: Effect of conventional anticancer agents against PANC-1 cells after 24 hours in NDM and DMEM. Data are expressed as the mean ± standard deviation, n=3. Abbreviations: NDM, nutrient-deprived medium; DMEM, Dulbecco’s Modified Eagle’s Medium; 5-FU, 5-fluorouracil; 2-DG, 2-deoxyglucose.

Article Snippet: Dulbecco’s Modified Eagle’s Medium. (DMEM) was purchased from Wako Pure Chemical (Osaka, Japan).

Techniques: Standard Deviation, Modification

Journal: Drug Design, Development and Therapy

Article Title: (+)-Grandifloracin, an antiausterity agent, induces autophagic PANC-1 pancreatic cancer cell death

doi: 10.2147/DDDT.S52168

Figure Lengend Snippet: Assessment of cytotoxicity of conventional anticancer agents against PANC-1 cells in Dulbecco’s Modified Eagle’s Medium. Data are expressed as the mean ± standard deviation, n=3. * P

Article Snippet: Dulbecco’s Modified Eagle’s Medium. (DMEM) was purchased from Wako Pure Chemical (Osaka, Japan).

Techniques: Modification, Standard Deviation

Journal: Drug Design, Development and Therapy

Article Title: (+)-Grandifloracin, an antiausterity agent, induces autophagic PANC-1 pancreatic cancer cell death

doi: 10.2147/DDDT.S52168

Figure Lengend Snippet: Effect of GF against Akt, mTOR, LC3A/B I, and LC3A/B II. Abbreviations: GF, (+)-grandifloracin; NDM, nutrient-deprived medium; DMEM, Dulbecco’s Modified Eagle’s Medium.

Article Snippet: Dulbecco’s Modified Eagle’s Medium. (DMEM) was purchased from Wako Pure Chemical (Osaka, Japan).

Techniques: Modification

Journal: Biomolecules

Article Title: High Levels of ROS Impair Lysosomal Acidity and Autophagy Flux in Glucose-Deprived Fibroblasts by Activating ATM and Erk Pathways

doi: 10.3390/biom10050761

Figure Lengend Snippet: Autophagy flux is impaired in glucose-deprived fibroblasts. ( A ) Human fibroblasts on cover slips were cultivated in normal (Glu(+)) or glucose-free Dulbecco’s modified Eagle’s medium (DMEM) (Glu(-)) for the indicated periods and processed for immunostaining for the LC3 protein (red). Nuclei were stained with 4′,6-diamidino-2-phenylindole (DAPI) (blue). Cells were examined by confocal microscopy. ( B ) Cells cultivated in glucose-free medium were examined by immunoblotting analysis for p62 sequestosome 1 (p62). ( C ) Change in LC3 protein level was determined by immunoblotting in cells cultivated in the absence of glucose for indicated times. The cells were either mock-treated or treated with 200-nM bafilomycin A1 (bafA1) or 50 μM chloroquine (CQ) for 6 h prior to collection for immunoblotting. Relative autophagic flux is shown in the bar graphs (below) of LC-II proteins levels quantitated by densitometry. The LC3-II level in the cells treated with bafA1 or CQ (+) was normalized by that in the cells untreated (-), and the ratio (R) is presented as an indication of the relative flux. ( D ) Cells transfected with plasmid that expresses tandem fluorescence LC3 (tfLC3) were cultivated in the absence of glucose for indicated periods or treated with 250-nM torin1 for 24 h. After fixing, tfLC3 fluorescence was visualized by confocal microscopy. In the right-most column, merged images (yellow) of the green and red fluorescence visualizes structures emitting both green and red fluorescence of GFP and RFP. All scale bars in microscopy indicate 5 μM.

Article Snippet: Cell Culture and Chemical Treatments Normal human fibroblasts isolated from healthy newborn foreskins and provided by Dr. Jin-Ho Chung (IRB No. H-1101-116-353 of the School of Medicine, Seoul National University, Korea) were maintained in Dulbecco’s modified Eagle’s medium (DMEM; LM001-11, Welgene, Daegu, Korea) supplemented with 10% fetal bovine serum (S-FBS-US-015, Serana, Bunbury, Australia) at 5% CO2 and 37 °C.

Techniques: Modification, Immunostaining, Staining, Confocal Microscopy, Transfection, Plasmid Preparation, Fluorescence, Microscopy