Fig. 5: ISRIB has concentration-dependent dichotomic effects on neuronal TyrRS levels.

a Lower doses of ISRIB increase TyrRS protein levels in neurons. Primary cortical neurons were treated with ISRIB (≤50 nM) for 8 hr and changes in the levels of TyrRS, PheRSβ, p-eEF2, eEF2, p-eIF2α, and eIF2α were detected and quantified by WB analysis using their respective antibodies. b Low dose ISRIB protects neurons against trans-RSV-mediated depletion of TyrRS. Primary cortical neurons were treated with trans-RSV (25 μM) alone or in combination with ISRIB (10 nM) for 8 hr, and changes in the levels of TyrRS were detected by WB. c High concentrations of ISRIB decrease TyrRS protein levels in cortical neurons. Primary rat cortical neurons were treated with ISRIB (100–500 nM) for 8 hr and changes in the levels of TyrRS, and PheRSβ were detected by WB analysis using their respective antibodies. All data represent mean ± SEM for n = 3 independent experiments with significance measured using two-way ANOVA with Dunnett’s test for multiple comparisons.