Fig 1: Increasing eEF2 levels enhance translation.A and B, Representative immunoblots for eEF2, Rpl7a and Rps6 (A) using lysates from HEK, immature and mature RCNs as well as their quantifications (B). Ribosomal proteins were used as loading controls (n = 6 biological replicates). C and D, Representative immunoblots against p-eEF2 and p-mTOR (C) using lysates from control and eEF2-HA transduced RCNs (18–22 DIV) as well as quantification (D). Ponceau served as loading control (n = 7 biological replicates for control, n = 6 biological replicates for eEF2-HA). E, polysome profiles (CHX treated) of control and eEF2-HA transduced RCNs (18–22 DIV). Shadows represent SEM of 3 biological replicates. F, steady-state P/M(0) upon eEF2-HA overexpression in RCNs (18–22 DIV, n = 3 biological replicates). p-values were calculated using One-way ANOVA with subsequent Tukey’s Multiple comparison test (B) and two-sided paired Student’s t-test (D and F). ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001. Dots represent biological replicates. Data are represented as mean ± SEM. n.s., not significant.