Fig. 7

Detection of ribosomal subunit interaction by PLA and two-color dSTORM. a Representative images of RPL24/puromycin-PLA signal detected in growth cones of Smn+/+;SMN2tgtg motoneurons. b PLA signal for RPL24/puromycin increases after 1-min BDNF stimulation (***P = 0.0001; n = 53–57 cells from 3 independent experiments). In control experiments, anisomycin treatment (a) decreased the RPL24/puromycin-PLA signal (****P < 0.0001; n = 18 cells from 1 experiment). c No PLA signal is detectable in the absence of primary antibodies to RPL24 or puromycin. Growth cones were outlined using GFP (see the method part for PLA assay). d and e Representative two-color dSTORM images labeling RPL24 and RPS6 (d) and Y10B and eEF2 (e) in growth cones of Smn+/+;SMN2tgtg neurons. Upon 1-min BDNF pulse, RPL24 and RPS6 as well as Y10B and eEF2 form clusters that tightly colocalize within growth cones. Arrows indicate positions where the RPL24/RPS6 and eEF2/Y10B signals are located in proximity of 50 nm or less. Data in b are presented in scatter dot plot; error bars represent mean ± SEM. Statistical analyses were done by one-way ANOVA with Dunn’s post-hoc test