Fig. 3.
Western blot analyses of proBDNF and mBDNF secretion induced by electrical stimulation at different frequencies. Rat hippocampal neurons (A) or tPA–/– neurons (B) transfected with BDNF construct shown in Fig. 1A were stimulated with the same number of electrical pulses (14,400) of HFS or LFS. Secretion of proBDNF and mBDNF was detected from culture medium (1 ml per well × 3 wells in a 12-well plate, with a plating density of 0.12 × 106 cells/well) by the method described in Fig. 1, using antibodies against the GFP tag and quantified by normalizing all data to average total BDNF signal under HFS condition (n = 3 independent experiments). ND, not detectable. Data are presented as mean ± SEM and analyzed by 2-way ANOVA followed by Bonferroni posthoc analysis. *, P < 0.01; **, P < 0.001.