Fig. 3.

Luciferase assays of uORF effects on protein and mRNA levels. (A) Experimental design of reporter constructs with and without uORFs is shown for example Mrpl11. (B–I) Normalized luciferase activity (B–E) and mRNA expression (F–I) are shown for reporter constructs that contain a uORF (red) or lack a uORF (gray) due to a mutation that disrupts the uORF start codon. The constructs contain 5′ UTRs from: 5 mouse genes chosen randomly (B and F), 10 mouse genes with proteomic and conservation signatures of functional uORFs (C and G), 5 human genes with polymorphic uORFs (D and H), and 5 human disease genes with uORF-altering mutations detected in patients (E and I). Error bars represent ±SE of ≥6 biological replicates (B–E) and ≥4 technical replicates (F–I). Asterisks indicate significant difference (P < 0.01).