Fig. 5
From: The non-coding RNA BC1 regulates experience-dependent structural plasticity and learning
Translation of PSD-95 mRNA is dysregulated in BC1 KO synaptoneurosomes. a left Representative western blotting showing incorporated puromycin in WT and BC1 KO cortical neurons. right Quantification of puromycin immunodetection normalized to Ponceau red staining (***P < 0.001 vs. WT, ### P < 0.001 vs. KO, one-way ANOVA, mean ± s.e.m., n = 9 WT, 16 KO (Cnt), 6 KO (4EGI) and 8 KO (CHX) mice). b Synaptoneurosomes from WT and BC1 KO mice were incubated with vehicle (Cnt) or 100 µM DHPG for 10 min in presence of L-AHA. Representative western blotting for all de novo-synthesized (biotin-positive) proteins detected using IRDye 800CW Streptavidin. Blots are different parts of the same gel (see Supplementary Fig. 6 for full scans). c left Representative western blotting showing the levels of ubiquitinated proteins in WT and KO synaptoneurosomes. right Quantification of ubiquitinated proteins normalized to Ponceau red staining (P = 0.4202; two-tailed t-test, mean ± s.e.m., n = 6 WT and 7 KO mice). n.s., not significant. d left Western blotting for de novo-synthesized (biotinylated) and total PSD-95, after immunoprecipitation (IP) with PSD-95 antibody. right Quantification of immunoblots; data are presented as the ratio (fold of WT-control) of biotinylated PSD-95 over total PSD-95 (*P < 0.05, two-way ANOVA, mean ± s.e.m., n = 6 WT and 5 KO mice). e Levels of BC1 RNA and of PSD-95 and αCaMKII mRNAs in cortical extracts from WT (white bars) and BC1 KO (black bars) mice. Values were normalized for HPRT and Gusb mRNA levels and expressed as fold of WT (mean ± s.e.m., n = 5 WT and 3 KO mice)
