|
|
|
|
|
|
|
|
|||||
| Home Help Feedback Subscriptions Archive Search Table of Contents | |||||
|
Fig. 5. vFLIP rescues MVECs from anoikis assayed by DNA fragmentation ELISA but does not rescue cells in the absence of growth factors. Human dermal MVECs were either not transduced or transduced with a lentiviral vector encoding GFP alone or vFLIP and GFP. 48 hours post transduction, cells were detached from the matrix by trypsinization and either replated on control wells (Attached –PH) or anoikis was induced by plating cells on polyHEMA (PH)-coated wells for 16 hours (Detached +PH). A subset of cells was washed free of medium and plated on control or PH-coated wells in the absence of any growth factors. (A) Apoptosis was measured by using a cell death detection ELISA kit that measures DNA fragmentation. Significant differences between the different samples are indicated on the graph. Differences were calculated using paired Student's t tests, in which P<0.05 was considered significant. *, P<0.05; **, P<0.01. (B) Cell metabolism was assessed using an MTT survival assay, which measures metabolic activity, as described in Materials and Methods. The results shown are the mean values from three independent experiments, with error bars indicating the standard deviation (s.d.) between the three experiments. Transduction efficiency was assessed by FACScan analysis of cells expressing GFP. 57% of MVECs infected with the vFLIP lentivirus were GFP positive, and 73% of MVECs infected with the GFP lentivirus were GFP positive.
|
