Extended Data Fig. 10: The rescuing effect of eIF5B in nutrient starvation induced SCARF.
From: Start codon-associated ribosomal frameshifting mediates nutrient stress adaptation
(a) A schematic image of 80S-eIF5B complex adopted from Wang et al, Nat Commun 2020. (b) Bar graphs show the HiBit-based SCARF reporter assays in cells transfected with wild type eIF5B or mutants. Both the Fluc (left panel) and HiBit (middle panel) signals were measured from cells transfected with SCARF reporters bearing a uORF start codon with or without the Kozak sequence context. Error bars, mean ± s.e.m.; n = 3. (c) Representative western blots of polyubiquitinated species in HEK293 cells with or without eIF5B overexpression before and after amino acid starvation. The experiment was independently repeated three times with similar results. (d) Representative western blots of exogenous eIF5B in transfected HEK293 cells. The experiment was independently repeated three times with similar results. (e) Representative western blots of ATF4 in starved HEK293 cells transfected with eIF5B wild type or mutants. The experiment was independently repeated three times with similar results.
