Journal: Hepatology (Baltimore, Md.)

Article Title: Apoptosis in human hepatoma cell lines by chemotherapeutic drugs via Fas-dependent and Fas-independent pathways.

doi: 10.1002/hep.510290102

Figure Lengend Snippet: FIG. 1. Expression of Fas in various hepatoma cell lines. (A) Fas messenger RNA was detected by reverse-transcriptase PCR using primers specific for Fas. (B) Surface expression of Fas was determined by flow cytometry using a mouse monoclonal antihuman Fas antibody (IgG, clone DX2). Open histogram with solid line, cells stained with secondary antibody alone; solid histogram, cells stained with anti-Fas antibody.

Article Snippet: Five different human hepatoma cell lines, Hep G2 (American Type Culture Collection [ATCC] HB8065, hepatitis B surface antigen [HBsAg] [2]), Hep 3B (ATCC HB 80640, HBsAg [1]), SK-Hep-1 (ATCC HTB 52, HBsAg [2]), PLC/PRF/5 (ATCC CRL 8024, HBsAg [1]), and Hep G2.2.15 (Hep G2 cell line transfected with a plasmid containing multiple copies of the hepatitis B virus genome)13-15 were maintained in minimum essential medium supplemented with 10% fetal bovine serum, 2 mmol/L glutamine, 100 IU/mL penicillin, and 100 μg/mL streptomycin in a 37°C humidified incubator under an atmosphere of 5% CO2 in air.

Techniques: Expressing, Reverse Transcription, Flow Cytometry, Staining

Journal: Hepatology (Baltimore, Md.)

Article Title: Apoptosis in human hepatoma cell lines by chemotherapeutic drugs via Fas-dependent and Fas-independent pathways.

doi: 10.1002/hep.510290102

Figure Lengend Snippet: FIG. 2. Effects of 5-FU or cisplatin on the growth inhibition of hepatoma cell lines. Adherent cells growing in 96-well plates (104 cells/well) were incubated with different concentrations of 5-FU or cisplatin (CP) for 48 hours (A), or with 25 µg/mL 5-FU or 15 µg/mL cisplatin for various time intervals (B). Cell growth was determined by MTT assay. Results are expressed as percent of cell growth of control at 48 hours (A) or 0 hours (B). The data shown are the mean from four independent experiments. Standard deviations are less than 10%.

Article Snippet: Five different human hepatoma cell lines, Hep G2 (American Type Culture Collection [ATCC] HB8065, hepatitis B surface antigen [HBsAg] [2]), Hep 3B (ATCC HB 80640, HBsAg [1]), SK-Hep-1 (ATCC HTB 52, HBsAg [2]), PLC/PRF/5 (ATCC CRL 8024, HBsAg [1]), and Hep G2.2.15 (Hep G2 cell line transfected with a plasmid containing multiple copies of the hepatitis B virus genome)13-15 were maintained in minimum essential medium supplemented with 10% fetal bovine serum, 2 mmol/L glutamine, 100 IU/mL penicillin, and 100 μg/mL streptomycin in a 37°C humidified incubator under an atmosphere of 5% CO2 in air.

Techniques: Inhibition, Incubation, MTT Assay, Control

Journal: Hepatology (Baltimore, Md.)

Article Title: Apoptosis in human hepatoma cell lines by chemotherapeutic drugs via Fas-dependent and Fas-independent pathways.

doi: 10.1002/hep.510290102

Figure Lengend Snippet: FIG. 3. Oligonucleosomal DNA fragmentation in hepatoma cell lines induced by 5-FU and cisplatin. Adherent cells growing in 100-mm dishes (2 3 106 cells/dish) were treated with 25 µg/ml 5-FU or 15 µg/mL cisplatin (CP) for 48 hours (A) or various time intervals (B). The fragmented DNA was extracted and subjected to agarose gel electrophoresis as described in Materials and Methods. m5marker.

Article Snippet: Five different human hepatoma cell lines, Hep G2 (American Type Culture Collection [ATCC] HB8065, hepatitis B surface antigen [HBsAg] [2]), Hep 3B (ATCC HB 80640, HBsAg [1]), SK-Hep-1 (ATCC HTB 52, HBsAg [2]), PLC/PRF/5 (ATCC CRL 8024, HBsAg [1]), and Hep G2.2.15 (Hep G2 cell line transfected with a plasmid containing multiple copies of the hepatitis B virus genome)13-15 were maintained in minimum essential medium supplemented with 10% fetal bovine serum, 2 mmol/L glutamine, 100 IU/mL penicillin, and 100 μg/mL streptomycin in a 37°C humidified incubator under an atmosphere of 5% CO2 in air.

Techniques: Agarose Gel Electrophoresis

Journal: Scientific Reports

Article Title: Molecular hydrogen suppresses activated Wnt/β-catenin signaling

doi: 10.1038/srep31986

Figure Lengend Snippet: (a) HCS-2/6 human chondrosarcoma cells were treated with control CM (Cont.), Wnt3a CM, or 2 μM BIO with 10% H 2 or 10% N 2 gas for 24 h. The Wnt/β-catenin signaling activity was measured by Topflash luciferase reporter assay ( n = 9). * P < 0.05 and ** P < 0.01 by Student’s t -test. (b) Human OAC cells were treated with control CM (Cont.), Wnt3a CM, or 2 μM BIO with 10% H 2 or 10% N 2 gas for 24 h. Representative Western blots are shown with densitometry of β-catenin/β-actin ( n = 4). * P < 0.05 by Student’s t -test. (c–g) Human OAC cells (clone 1) were treated with control CM (Cont.), Wnt3a CM, or 2 μM BIO with 10% H 2 or 10% N 2 gas for 24 h. Expression of AXIN2 ( n = 3) (c) , SOX9 ( n = 6) (d) , MMP3 ( n = 6) (e) , COL2A1 ( n = 3) (f) , and ACAN ( n = 3) (g) were quantified by qRT-PCR. * P < 0.05 by Student’s t -test. (h) Differentiated ATDC5 cells were treated with control CM (Cont.), Wnt3a CM, or 2 μM BIO with 10% H 2 or 10% N 2 gas for 48 h. Proteoglycans were stained with Alcian blue (upper panel) and quantified by measuring the optical intensity at 630 nm of the cell lysates ( n = 9 for Cont.-treated and BIO-treated cells, n = 6 for Wnt3a-treated cells). * P < 0.05 by Student’s t-test. Non-significant P values less than 0.10 are indicated above each pair.

Article Snippet: HepG2 and ATDC5 cells were from RIKEN BioResource Center.

Techniques: Control, Activity Assay, Luciferase, Reporter Assay, Western Blot, Expressing, Quantitative RT-PCR, Staining