Fig. 6

BDNF-induced formation of ribosomal complexes can be visualized by proximity ligation assay. a Representative images show PLA signal from RPL24 and RPS6 proteins in growth cones of Smn+/+;SMN2tgtg and Smn−/−;SMN2tgtg motoneurons following 1-min BDNF stimulation. b Increased PLA signal is detectable in growth cones of BDNF-stimulated Smn+/+;SMN2tgtg (****P < 0.0001; n = 65–69 cells from 4 independent experiments), but not in Smn−/−;SMN2tgtg neurons (n.s., P = 0.9315; n = 39 cells from 3 independent experiments). As a negative control, Smn+/+;SMN2tgtg neurons were pretreated with puromycin before and during the BDNF pulse (a). Only a low PLA signal is detectable (n.s., P = 0.6882; n = 13 cells from 1 experiment). c Omitting either the RPL24 or the RPS6 primary antibody  abolishes the PLA signal. Growth cones were outlined using GFP (see the method part for PLA assay). Data are presented in scatter dot plot; error bars represent mean ± SEM. Statistical analyses were done by Two-way ANOVA with Tukey multiple comparison post-hoc test