Figure 3.
BDNF gene transfer prevents apoptotic cell death of FXN-deficient neurons in culture. Primary cortical neurons were transduced with lentivectors (shRNA-sc or shRNA-37) or were left untransduced as a mock control (m), in the presence of HSV-1 amplicon vectors containing the cDNA of BDNF (HSV-BDNF) or the cDNA of β-galactosidase (HSV-LacZ) as control. (a) Representative western-blot analysis of FXN, cleaved caspase-3, β-galactosidase, and BDNF levels at 72 hours upon cotransduction. (b) Quantification of cleaved caspase-3 levels in FXN-deficient neurons upon cotransduction reveals a decrease of caspase-3 activation when BDNF was overexpressed. (c) Cell viability of cultured neurons at 72 hours upon cotransduction as assessed by the calcein/propidium iodide assay shows significant differences between FXN-deficient neurons with or without HSV-BDNF. (d) Cell metabolic activity of cultured neurons at 72 hours upon cotransduction was assessed by MTS reduction and also shows significant differences between FXN-deficient neurons with or without HSV-BDNF. Data represent mean values ± SEM from three independent experiments, *P ≤ 0.05, **P ≤ 0.005.