Journal: PLoS ONE

Article Title: Epigenetic Upregulation of Endogenous VEGF-A Reduces Myocardial Infarct Size in Mice

doi: 10.1371/journal.pone.0089979

Figure Lengend Snippet: (a) ELISA analysis of VEGF-A protein from transduced hearts, (b) ELISA assay from growth medium of C166 cells transduced with LV-451 and corresponding single stranded vectors using MOI 10, 7 days time point. (c) RT-PCR analysis of VEGF-A mRNA levels. C166 cells were transduced with LV-451 and corresponding single stranded vectors using MOI 10, 11 days time point. (d) qChIP assay of C166 cells using antibodies against H3K4me2. Cells were transduced with LV-451 and corresponding single stranded vectors using MOI 10, 11 days timepoint. All results are shown as mean ± SD.

Article Snippet: C166 (ATCC: CRL-2583) cells were cultured in DMEM containing fetal bovine serum (FBS, 10%), 100 units/ml penicillin and 100 µg/ml streptomycin.

Techniques: Enzyme-linked Immunosorbent Assay, Transduction, Reverse Transcription Polymerase Chain Reaction

Journal: PLoS ONE

Article Title: Epigenetic Upregulation of Endogenous VEGF-A Reduces Myocardial Infarct Size in Mice

doi: 10.1371/journal.pone.0089979

Figure Lengend Snippet: C166 cells were subjected to RNA-FISH analysis with LV-451 or VEGF mRNA probes. (a) Confocal microscopy images of LV-451 transduced (MOI 10) cells 72 h post transduction. Distribution of LV-451 RNA (green) and VEGF-A mRNA (red) probe binding induced signals is shown. Nuclei were visualized with DAPI (grey). Scale bars, 5 µm. (b) Quantification of LV-451 RNA or VEGF-A mRNA RNA-FISH signal spots detected in LV-451 transduced (MOI 4, 40, 200) cells at 72 h post transduction and in nontransduced control cells. The amount of signal was calculated in the nucleus (white), the cytosol (grey) and whole cell (black). Error bars = SD. (c) Nucleus size in response to LV transduction. CTRL sample is nontransduced C166 cells and LV-451 is C166 cells transduced with LV-451 vector.

Article Snippet: C166 (ATCC: CRL-2583) cells were cultured in DMEM containing fetal bovine serum (FBS, 10%), 100 units/ml penicillin and 100 µg/ml streptomycin.

Techniques: Confocal Microscopy, Transduction, Binding Assay, Plasmid Preparation

Journal: PLoS ONE

Article Title: Epigenetic Upregulation of Endogenous VEGF-A Reduces Myocardial Infarct Size in Mice

doi: 10.1371/journal.pone.0089979

Figure Lengend Snippet: (a) RT-PCR analysis for different VEGF-A isoforms. The expression levels for different isoforms were studied using primers specific to each isoform. Total VEGF-A protein level was measured with ELISA. (b) Reversing DNA methylation with 5-Azacytidine treatment induces responses in MS1 cells but erases responses in C166 cells. Cells were treated with 1 µM 5-Azacytidine, transduced with different vectors on day 3 and samples were collected on day 8. qRT-PCR analysis of VEGF-A and B-actin mRNA levels in MS1 cells and C166 cells. (c) qChIP assay in MS1 cells using antibody against H3K27me3. (d) The VEGF-A gene promoter in C166 cells was also analyzed for basal DNA methylation levels without 5-Azacytidine treatment using MeDIP. Cells were transduced with different vectors using MOI 10, 10 days timepoint. (e) RT-PCR analysis of VEGF-A mRNA levels after C166 cells were transfected with siRNA oligos. Results are calculated in reference to housekeeping gene ACTB and control oligo. (f) CBP-CREB interaction inhibitor (7.5 µM) abolishes the upregulation of VEGF-A by LV-451 in C166 cells. For all results, mean ± SD shown.

Article Snippet: C166 (ATCC: CRL-2583) cells were cultured in DMEM containing fetal bovine serum (FBS, 10%), 100 units/ml penicillin and 100 µg/ml streptomycin.

Techniques: Reverse Transcription Polymerase Chain Reaction, Expressing, Enzyme-linked Immunosorbent Assay, DNA Methylation Assay, Transduction, Quantitative RT-PCR, Methylated DNA Immunoprecipitation, Transfection

Journal: Pharmaceutics

Article Title: Chitosan-Coated Liposome Formulations for Encapsulation of Ciprofloxacin and Etoposide

doi: 10.3390/pharmaceutics16081036

Figure Lengend Snippet: Quantification of cellular metabolic activity via the AlamarBlue technique in cells of the C166-GFP line.

Article Snippet: The cytotoxicity of etoposide-containing liposomes and CS-coated liposomes was evaluated in C166-GFP mouse endothelial cell line (ATCC CRL2583™, Manassas, VA USA) through an analysis of their metabolic activity.

Techniques: Activity Assay