Journal: Journal of Cellular and Molecular Medicine

Article Title: 5′AMP-activated protein kinase α deficiency enhances stress-induced apoptosis in BHK and PC12 cells

doi: 10.1111/j.1582-4934.2007.00023.x

Figure Lengend Snippet: Apoptosis induction on treatment of HSVTK + BHK cells with PCV and GCV. Western blot analysis shows high levels of cleaved Caspase-3 at 48 hrs and 70 hrs of treatment with PCV and GCV. PCV and GCV treatment also induces cleavage of the Caspase-3 substrate, PARP, which is not observed on treatment with ACV. (NT = non-treated control.)

Article Snippet: GCV, PCV, and ACV were obtained from GlaxoSmithkline Research and Development, Stevenage, UK.

Techniques: Western Blot, Control

Journal: Journal of Cellular and Molecular Medicine

Article Title: 5′AMP-activated protein kinase α deficiency enhances stress-induced apoptosis in BHK and PC12 cells

doi: 10.1111/j.1582-4934.2007.00023.x

Figure Lengend Snippet: Western blot confirming enhanced phosphorylation of AMPKα in ( A ) PC12 cells exposed to low (3 mM) and glucose-free medium (11.1 mM glucose = normal culture conditions) and ( B ) HSVTK + BHK cells treated with ACV, PCV or GCV.

Article Snippet: GCV, PCV, and ACV were obtained from GlaxoSmithkline Research and Development, Stevenage, UK.

Techniques: Western Blot, Phospho-proteomics

Journal: Journal of Cellular and Molecular Medicine

Article Title: 5′AMP-activated protein kinase α deficiency enhances stress-induced apoptosis in BHK and PC12 cells

doi: 10.1111/j.1582-4934.2007.00023.x

Figure Lengend Snippet: ( A ) Effect of AMPK downregulation on the extent of guanosine nucleoside analogue-induced death in shRNA-transformed HSVTK + BHK cells at (i) 24 hrs, (ii) 48 hrs and (iii) 72 hrs of treatment. Cell death was measured by propidium iodide staining and flow cytometry analysis. ‘Dead’ cells were counted as propidium iodide-positive events scoring above the 102 value on the FL2 scale. Data are averaged from n = 3 clones from the respective shRNA-expressing groups (green = p Silencer /negative control shRNA, blue = p Silencer /AMPKα 1 shRNA, red = p Silencer / AMPKα 2 shRNA-transformed cells). Asterisk indicates significant difference between levels of cell death in the negative control shRNA and AMPKα shRNAexpressing cells, treated with 10 μM PCV or GCV (p < 0.05, Student's t-test). ( B ) Flow cytometry analysis of propidium iodide-stained p Silencer /negative control (i), AMPKα 1 (ii) or AMPKα 2 (iii) shRNA-transformed PC12 cells, cultured for 4 days in medium with (green plots) or without (red plots) glucose. Enhanced cell death is evident in the p Silencer /AMPKαshRNAtransformed cells. ( C ) Flow cytometry analysis of propidium iodide-stained, non-transformed PC12 cells, cultured for 4 days without compound C (i), with 0.1 μM compound C (ii) or with 1μM compound C (iii), in medium with (green plots) or without (red plots) glucose. Glucose deprivation-induced death is enhanced in the compound C-treated cells.

Article Snippet: GCV, PCV, and ACV were obtained from GlaxoSmithkline Research and Development, Stevenage, UK.

Techniques: shRNA, Transformation Assay, Staining, Flow Cytometry, Clone Assay, Expressing, Negative Control, Cell Culture