Figure 5. Requirement of PDK1 for anti-apoptotic activity of BDNF in trophic-deprived neurons.
Neurons were co-transfected with shGFP or an equimolar mix of shPDK1-1 and shPDK1-2 (shPDK1) together with the β-gal expression plasmid, pON260 (1 + 0.2 µg plasmid DNA/5×105 neurons, respectively). After 24 hours, neurons were sham-treated or trophic deprived in the presence of 0 or 10 ng/ml BDNF for the next 24 hours. A, Representative photomicrographs illustrating morphology of the transfected neurons, which were identified by β-gal immunostaining. Counterstaining with Hoechst 33258 revealed apoptotic alterations including condensation and fragmentation of the chromatin (arrows). Arrowheads point β-gal-positive cells without signs of apoptosis. B, The knock down of PDK1 reduced BDNF protection against TD-induced apoptosis. The individual shPDK1 plasmids affected BDNF survival similarly to their mix (data not shown). Averages of triplicate determinations from 3 independent experiments ±SEM are presented; *, p<0.05; ***, p<0.001.