Fig. 2.

Cisplatin increases both BDNF protein production and mRNA translational efficiency. (A) ELISA detection of total BDNF protein level in 4.5 DIV differentiated SK-N-BE cells after cisplatin treatment, at different time points: untreated control (0), 1, 3, 6, 12 and 24 h (N=4, each in duplicate). (B) Evaluation of mature BDNF protein production through densitometric analysis of western blot, in the same conditions as described in panel A, but limited to time point treatment of 6 and 24 h; BDNF signal is normalized to tubulin and given as percentage of variation with respect to control. Lower panel: representative western blot of mature BDNF and tubulin (N=3). (C) Translational efficiency of different BDNF isoforms under cisplatin cytotoxic stress, at 6 and 24 h time points. The isoforms tested are as follows: exon1-Fluc-3′UTR long (Ex1-F-3′L), exon2c-Fluc-3′UTR long (Ex2c-F-3′L), exon4-Fluc-3′UTR long (Ex4-F-3′L) and exon6-Fluc-3′UTR long (Ex6-F-3′L). Values from each isoform are normalized for transfection efficiency over a control vector (Rluc) and given as fold increase on basal condition (0); N=3, each in duplicate; RLU: relative luciferase unit. In all panels, data are represented as mean±s.e.m. *P<0.05, **P<0.01, ***P<0.001; one-way ANOVA versus untreated control.