Fig. 6: Rpl10a ^LOF/LOF embryos exhibit reduced translation of Wnt signaling components.

a (Left) Polysome traces of wild-type, Rpl10a ^LOF/+, and Rpl10a ^LOF/LOF whole E13.5 embryo extracts show similar distributions of polysomes, monosomes, and free subunits. (Right) Western blots of protein precipitated from each sucrose-gradient fraction show the extended RPL10A/uL1 protein is incorporated into polysomes equivalently to wild-type. b (Left) Fluorescence image of E9.5 Meox1 ^Cre; Ai9 embryo illustrating the presomitic/somitic mesoderm tdTomato reporter expression. (Right) OP-Puromycin incorporation rates in Meox1 ^Cre; Ai9; Rpl10a ^LOF/LOF E9.5 presomitic/somitic mesoderm (tdTom+) and in the rest of the embryo (tdTom−) (n = 3). Values are normalized to OP-Puro incorporation rates of TdTom+ and TdTom– cells of Meox1 ^Cre; Ai9; Rpl10a ^LOF/+ E9.5 control littermates. Data shown are mean +/− SEM, and significance was calculated using Student’s t test. c Comparison of RNA-seq and ribosome profiling for whole E8.5 Rpl10a ^LOF/LOF and wild-type embryos (n = 3 each). Genes changing significantly (false discovery rate (FDR) < 0.1) only in mRNA abundance are blue; genes changing significantly (FDR < 0.1) only in ribosome occupancy are red; genes changing significantly in both datasets are purple. d Selected Wnt signaling gene sets that were significantly (FDR < 0.1) enriched in CAMERA gene set enrichment analysis of genes with altered translation efficiency in Rpl10a ^LOF/LOF embryos compared to wild-type. All gene sets shown are translationally downregulated. Node size = gene set size; edge size = gene set overlap. e Genes from the “Wnt signaling pathway, planar cell polarity pathway” set with statistically significant changes in ribosome-protected footprints (RPF) but not in total RNA-Seq (RNA). f E9.5 wild-type (n = 2) and Rpl10a ^LOF/LOF (n = 4) gradient RT-qPCR for Vangl2 showing the fraction of the total mRNA found in each of the five fractions of the 25–50% sucrose gradient demarcated in the schematic. Data shown are mean +/− SEM. Vangl2 is significantly decreased in Rpl10a ^LOF/LOF medium and heavy polysomes relative to wild-type (Student’s t test P values = 0.04 and 0.008, respectively). *P value < 0.05; **P value < 0.01. Source data are provided as a Source Data file.