Fig. 7
From: Critical and differential roles of eIF4A1 and eIF4A2 in B-cell development and function
eIF4A1 controls the translation of Gins4 via 5’UTR. A Experimental outline of the RNAseq and LFQ-MS experiments. B KEGG pathway enrichment of downregulated proteins identified by LFQ-MS and downregulated mRNAs quantified by RNA-seq. C Venn diagram analysis of differentially expressed genes (DEGs) identified via RNA-seq (RNA-DEGs) and LFQ-MS (MS-DEGs). Cutoff, absolute Log2FoldChange greater than 0.5. D A total of 248 genes from C with only altered protein expression were analyzed via Metascape for GO enrichment, and the top ten terms were plotted. E Immunoblot analysis of eIF4A1, Gins1, and Gins4 in Eif4a1fl/fl and Eif4a1fl/fl;CD19Cre B cells stimulated with LPS for the indicated amounts of time. F RNA structures of Gins4 and Actb 5’UTRs were predicted via RNAfold. MFE, minimum free energy. G Distribution of Actb and Gins4 mRNAs in the sucrose gradient fractions of Eif4a1fl/fl and Eif4a1fl/fl;CD19Cre B cells stimulated with LPS for 24 hours. H Graphical outline of the luciferase reporter gene. I Immunoblot analysis of eIF4A1 protein expression in HEK293T cells transfected with scramble or Eif4a1 siRNA (left panel). Luciferase assay of the reporter gene harboring the Actb or Gin4 5’UTR H in HEK293T cells transfected with scramble or Eif4a1 siRNA (right panel). The error bars represent the standard errors of the means (SEMs). ns, not significant; *p < 0.05, **p < 0.01 and ***p < 0.001. The data shown are representative of three independent experiments
