Fig. 6: EIF3B promotes global translation downstream of PRDM10.
From: Global translation during early development depends on the essential transcription factor PRDM10
a Immunoblotting detection of FLAG-tagged EIF3B protein expression in Prdm10F/F; CreERT2 mESCs transformed with pJ549-Eif3b or vector. b qRT-PCR validation of Eif3b transcript in Prdm10F/F; CreERT2 mESCs transformed with pJ549-Eif3b or vector, analyzed 6 days post-induction. CDS-specific primers were used to detect both endogenous and overexpressed Eif3b transcript; n = 3 biological replicates. c Growth curve analysis of Prdm10F/F and Prdm10Δ/Δ mESCs in the presence of empty vector or exogenous Eif3b, showing partial phenotypic rescue by Eif3b overexpression. Cells were passaged at constant density every 2 days and counted daily up to Day 8 post-induction; n = 3. Representative data shown from one of three independent experiments. d Average population doubling times over the course of the growth assay for Prdm10F/F and Prdm10Δ/Δ mESCs expressing vector control or exogenous Eif3b. Each point represents data from one independent experiment; n = 3. e Representative polysome profiles of Prdm10F/F and Prdm10Δ/Δ mESCs analyzed 5 days post-induction (left). A significantly reduced polysome:monosome (P/M) ratio indicates impaired global translation in Prdm10Δ/Δ mESCs (right); n = 5 biological replicates, across two independent experiments. f Representative polysome profiles of Prdm10F/F and Prdm10Δ/Δ mESCs expressing Eif3b or vector, analyzed at Day 5 post-induction (left). Eif3b overexpression restores global translation in Prdm10Δ/Δ mESCs to levels comparable to Prdm10F/F controls, as quantified by P/M ratio (right); n = 5 biological replicates across two independent experiments. Gene expression presented relative to Ubb (b). Data presented as mean ± s.d.; **P < 0.01, ****P < 0.0001, n.s., not significant; one-way ANOVA with Tukey’s multiple comparisons test (b, d and f); two-way ANOVA with Tukey’s multiple comparisons test (c).
