sh-sy5y cells  (CLS Cell Lines Service GmbH)

Journal: PLoS ONE

Article Title: A novel understanding of postoperative complications: In vitro study of the impact of propofol on epigenetic modifications in cholinergic genes

doi: 10.1371/journal.pone.0217269

Figure Lengend Snippet: ( a ) We could not detect any significantly modified methylation in the promoter of the ACHE gene (area -1703/-1559) and BCHE gene (intron 2), apart from a decreased methylation in the promoter of the CHRNA7 gene (area -1185/-1010; p = 0.011) after stimulating SH-SY5Y cells with 25 μg/ml propofol for 2 h. Methylation of CHRNA7 recovered after 4h to almost unstimulated levels. ( b ) The artificial de-methylation of the epigenome of SH-SY5Y cells using 5-aza-2’ deoxycytidine (ADC, 50 μM) resulted in a visible decrease of overall methylation as measured in the three promoter regions. ( c ) This de-methylation reduced the expression of CHRNA7 mRNA (p = 0.034) but did not affect ACHE or BCHE significantly. ( d ) Propofol did not change the expression of proinflammatory (TNFα and IL-6) and anti-inflammatory cytokines (IL-10). Error bars depict 2 x SE.

Article Snippet: The human neuroblastoma cells SH-SY5Y (origin: Cell Lines Service, CLS, Eppelheim Germany, SH-SY5Y item number: 300154) were cultured at 37°C and 5% CO 2 in Dulbecco’s modified Eagle medium (DMEM; Gibco, Darmstadt, Germany) with 10% foetal calf serum (FCS; Gibco, Darmstadt, Germany) and 1% penicillin/streptomycin (Penstrep; Gibco, Darmstadt, Germany).

Techniques: Modification, Methylation, Expressing

Journal: PLoS ONE

Article Title: A novel understanding of postoperative complications: In vitro study of the impact of propofol on epigenetic modifications in cholinergic genes

doi: 10.1371/journal.pone.0217269

Figure Lengend Snippet: ( a ) The tri-methylation of H3 K27 in SH-SY5Y cells was significantly decreased by propofol [25 μg/ml], while TNFα [10 ng/ml] did not show any changes after 24 h. ( b ) ChIP assays showed that the promoter regions of ACHE , BCHE and CHRNA7 bind to tri-methylated H3 K27. For the negative control (1) rabbit IgG was used instead of the specific tri met H3 K27 antibody (2), and the positive control contained an antibody against RNA polymerase and primers for GAPDH. ( c ) Propofol increased the expression of the histone de-acetylating HDAC1 approximately 1.5-fold. ( d ) The expression of KDM2A was increased more than 3-fold after 4h, and ( e ) the expression of DMNT3B was slightly increased over the first 4 hours. Error bars depict 2 x SE.

Article Snippet: The human neuroblastoma cells SH-SY5Y (origin: Cell Lines Service, CLS, Eppelheim Germany, SH-SY5Y item number: 300154) were cultured at 37°C and 5% CO 2 in Dulbecco’s modified Eagle medium (DMEM; Gibco, Darmstadt, Germany) with 10% foetal calf serum (FCS; Gibco, Darmstadt, Germany) and 1% penicillin/streptomycin (Penstrep; Gibco, Darmstadt, Germany).

Techniques: Methylation, Negative Control, Positive Control, Expressing