Figure 6. Translation attenuation in response to tRNA synthetase inhibitors is independent of the ISR.

1. Newly synthesized proteins pulse‐labeled with ³⁵S‐methionine for 10 min in HeLa cells pre‐treated with indicated compounds for 2.5 h, except (E) (Bottom) Coomassie‐stained gel.
2. Newly synthesized proteins in HeLa cells treated with 2.5 μg/ml Tm or 50 μg/ml cycloheximide (CHX).
3. Newly synthesized proteins in 4KO or 4KO+GCN2 MEF cells treated with 12.5 or 200 nM HF.
4. Newly synthesized proteins in eIF2α^S/S and eIF2α^A/A MEF cells treated with 12.5 or 200 nM HF.
5. Newly synthesized proteins in eIF2α^S/S and eIF2α^A/A MEF cells treated with indicated concentrations of Tm for 2.5 or 5 h.
6. Newly synthesized proteins in HeLa cells untreated or treated with GCN2 siRNA 48 h before treatment with indicated concentrations of HF.
7. Newly synthesized proteins in HeLa cells treated with indicated concentrations of HF with or without proline supplementation (10 mM).
8. Representative immunoblots of indicated proteins in lysates from HeLa cells after treatment with indicated concentrations of HF with or without proline supplementation (10 mM).
9. Newly synthesized proteins in HeLa cells untreated or treated with GCN2 siRNA 48 h before treatment with indicated concentrations of histidinol (His) or borrelidin (Bor).

Data information: Quantification of experiments such as the ones shown in (C), (D) and (I) are presented in Figs EV3 and EV4.