Fig. 8: Pharmacological targeting of eIF6 activity reduces steatosis in murine hepatocytes.

a Model of the translational regulation of lipogenic TFs driven by eIF6. eIF6 regulates the translation of C/EBPβ mRNA, which contains uORF in its 5’UTR, through re-initiation. eIF6 has an anti-association property: it binds the 60 S ribosomal subunit at the stop codon of the uORF, leading to 60 S release from the 40 S. 40 S can then resume scanning to the main ORF, which encodes for the transcription factor (TOP). eIFsixty compounds prevent eIF6 binding to the 60 S at the stop codon of the uORF. In this case, the inactive 80 S complex is stalled and the translation of the downstream ORF is inhibited (BOTTOM). b Bright-field images of AML12 treated with 2 mM palmitate and with eIFsixty-i compounds. Cells treated with palmitate have increased accumulation of lipid droplets, which is reduced upon eIFsixty-1 and eIFsixty-6 inhibitors treatment. eIFneg is a negative control, an inactive compound of a similar structure. Scale bar = 50 µm. c Quantification of Oil Red O content in palmitate-treated AML12 cells. N = 5/condition. Data are represented as means ± SD. Two-tailed t test. d Quantification of mRNA levels in heavy, light, and subpolysomes of AML12 cells after administration of palmitate and eIFsixty-i compounds. eIFneg was used as a negative control. Only eIFsixty-1 and eIFsixty-6 induce a shift from the polysomal to the subpolysomal peak of eIF6 target C/EBPβ mRNA. The same treatment shows the opposite effect on YY1 mRNA. Real-time PCR analysis was performed on n = 3 independent experiments. Stacked bar charts represent the quantification of selected mRNA levels in heavy, light, and subpolysomes. Data are provided as a Source Data File.