m 5 aza  (Toronto Research Chemicals)

Journal: Cell Death & Disease

Article Title: A metabolic synthetic lethal strategy with arginine deprivation and chloroquine leads to cell death in ASS1-deficient sarcomas

doi: 10.1038/cddis.2016.232

Figure Lengend Snippet: ADI-PEG20 treatment inhibits tumor growth until ASS1 re-expression confers resistance. ( a ) Flow cytometry analysis of cell death induction, as measured by percentage of cells staining positive for PI, upon treatment with ADI-PEG20. As a single agent, ADI-PEG20 does not induce cell death in these sarcoma cell lines. ( N =3). Data represented as mean±S.D. ( b ) Tumor growth of ASS1 low SK-LMS-1 cells xenografted into nude mice with or without ADI-PEG20 treatment. Significant tumor growth inhibition was observed when mice were treated with ADI-PEG20 as compared with tumor growth in PBS-treated mice. Shown below is a western blot of five tumor lysates from five mice after ADI-PEG20 treatment showing re-expression of ASS1 in tumors, which had gained resistance to ADI-PEG20 mediated growth inhibition. ( N =5 mice per arm). Data represented as mean tumor volume±S.E.M. ( c ) Re-expression of ASS1 in SK-LMS-1 cells after treatment with 1 μ g/ml ADI-PEG20, 5 μ M 5-aza-dC or both, as compared with untreated cells. After 48 h of combination treatment, ASS1 expression levels have significantly increased above wild-type conditions, as well as either drug individually. ( N =3)

Article Snippet: Chloroquine (used at 10 and 20 μ M), 5-aza-dC (5 μ M), necrostatin (10 μ M) and ZVAD-FMK (100 μ M) were obtained from Sigma Aldrich (St. Louis, MO, USA).

Techniques: Expressing, Flow Cytometry, Staining, Inhibition, Western Blot

Journal: The Journal of Biological Chemistry

Article Title: The Role of Embryonic Stem Cell-expressed RAS (ERAS) in the Maintenance of Quiescent Hepatic Stellate Cells *

doi: 10.1074/jbc.M115.700088

Figure Lengend Snippet: DNA methylation analyses of ERas in quiescent and activated primary HSCs. A, results of genome-wide DNA methylation analysis of quiescent HSCs (d0)and early activated HSCs (d3) at the ERas promoter region. DNA methylation of individual CpG dinucleotides is depicted in percent (methylated CpG/total numbers of CpG) and displayed with a color code from red (0%) to light green (100% DNA methylation). B, ERas promoter methylation analyzed by direct bisulfite sequencing exhibited a significant increase of DNA methylation during HSCs activation (n = 3–5, t test; *, p < 0.05). C, HSC (day 3) were treated with 10 μm 5-aza-2′-deoxycytidine (5-AZA) and/or 5 μm suberoylanilide hydroxamic acid (SAHA) for 4 days, and the expression of ERas was analyzed with qPCR at day 8. Error bars, S.E.

Article Snippet: Primary rat HSCs at day 3 were treated with 10 μ m 5-aza-2′-deoxycytidine (5-AZA) (Decitabine, Sigma catalog no. A3656), a specific DNA methyltransferase inhibitor, for 4 successive days.

Techniques: DNA Methylation Assay, Genome Wide, Methylation, Methylation Sequencing, Activation Assay, Expressing