Extended Data Fig. 10: Translational upregulation of dendritic downstream ORFs by eIF4G2 is mediated by its local calcium-influx-induced phosphorylation.
From: Neuronal activity rapidly reprograms dendritic translation via eIF4G2:uORF binding
a, CDF plots of dendritic translation of eIF4G2-bound transcripts that harbor uORFs in wild type and eIF4G2 knockdown conditions. Significance was calculated by the two-sided Wilcoxon signed-rank test. b, Dendritic enrichment (log2) of eIF4G2 is shown using the resting and depolarized PL-MS data (n = 5, values from PL-MS data). Significance was calculated using the two-tailed, unpaired Student’s t-test. Box plot whiskers extend from min to max, with the center line at median. c, Nascent translation of Nsun3 and Slc25a19, examples of nuclear-encoded mitochondria-related RNAs, is shown by PLA (red) in the presence (EGTA-) and upon depletion (EGTA+) of calcium in resting and KCl-depolarized neurons (n = 2). DAPI (blue, marker for nuclei). d, Shown are the Flag and β-Actin western blots that are quantified in Fig. 5g. eIF4G2 western blot shows endogenous eIF4G2 (lower bands), which is reduced by siRNA knockdowns (Kd), as well as the localized wild type and phospho-mutant eIF4G2 variants (upper bands, higher molecular weight due to the inclusion of dendritic localization signals). Fold changes are shown as in Fig. 5g but here significance was calculated with respect to the knockdown, using the two-tailed, paired Student’s t-test (n = 3). (c,d) All data are presented as mean ±s.d. n indicates the number of biologically independent samples.
