By evidence of a DNA damage response consistent with telomere uncapping

Intramuscular transplantation and non-invasive bioluminescence NS-018 imaging was 1608125-21-8 biological activity performed under 1�C4 1L O2/min isoflurane inhalation. Euthanasia was performed under isoflurane inhalation followed by cervical dislocation. Primary myoblasts were isolated from skeletal muscle of 2 month old C57BL/6 and Luciferase x EGFP mice as described previously , plated on tissue culture plates coated with collagen and maintained in growth media . To expose cells to normoxic or hypoxic culture conditions, cultures were placed in an airtight modular hypoxia chamber adjusted to the indicated oxygen concentration. The EMD kinase inhibitor library was screened for their capability to protect cells from hypoxia- induced myoblast cell death/growth arrest. The cells were plated at 1500 cells/well in 384-well plates in growth media. At least 4 hours after cell seeding, 244 kinase inhibitors were dispensed into the cells-seeded plates at 1 ��Mfinal concentration using Echo liquid handler . The cells were cultured under hypoxic environment created by the infusion of a gas mixture of 95 of N2 and 5 of CO2 into an airtight modular hypoxia chamber for 5 days. Two independent screens were performed with duplicates each run. Muscle tissues were prepared for histology as previously described . Cells and muscle sections were fixed with 1.5 PFA, permeabilised in 0.3 Triton and blocked in 20 goat serum. Incubation with the primary antibodies was performed overnight at 4. The antibodies used are: rabbit anti-GFP , rat anti-laminin , rabbit anti-hypoxyprobe , rabbit anti-HIF-1�� , rabbit anti-cCasp3 and Alexa-conjugated secondary antibodies . Images of cell cultures as well as muscle transverse sections were acquired using an inverted epifluorescent microscope , 10x objective lens, CCD SPOT RT camera and SPOT imaging software . Fluorescent intensities of selected immunofluorescent regions were measured as mean gray values . All images were composed, edited and modifications applied to the whole image using Photoshop CS6 . Pathway analysis was obtained by combining two datasets containing drug-target information , one datasets containing protein-protein interaction