Fig. 6

ROS induction reduces protein translation in the cytosol. a–e Incorporation of [³⁵S]-labelled amino acids in yeast or mammalian cells. Total cell extracts were separated by SDS-PAGE and analysed by autoradiography or immunodecorated with specific antibodies. a Wild-type (BY4741) yeast cells were grown on fermentative medium and treated for 30 min with H₂O₂ as indicated. b HEK 293 cells were treated for 2 h with H₂O₂ as indicated. c Yeast cells were grown on respiratory medium at 19 °C and shifted for 2 h to restrictive temperature (37 °C). Incorporation of [³⁵S]-labelled amino acids in wild-type yeast and mia40-4int mutant was done for 1 h before collection of cells. d, e Wild-type (BY4741) yeast cells and cells with deletions of ribosomal genes (encoding for proteins that according to the proteomics data were significantly more oxidised in mia40-4int and upon H₂O₂ treatment (d) or significantly more oxidised upon H₂O₂ treatment only (e)) were grown on fermentative medium and treated for 30 min with H₂O₂ (upper panel). Quantification of protein synthesis. Mean ± SEM, n = 3 (lower panel). WT, wild-type