Fig. 5

Mechanistic validation of the TrioSig gene set in cell culture. A Diagram of the ISR and NRF2 pathways. B Levels of peIF2α, total eIF2α, NRF2, ATF4, and actin after treatment of HT22 cells with 5 mM glutamate, 500 nM erastin, and 250 nM RSL3 at various timepoints were assessed by western blotting and quantified (n = 3–4/condition). C Levels of NRF2, ATF4, and actin after knockdown with control (Ct), Nrf2, or Atf4 siRNAs (n = 7/condition). D Survival of HT22 cells exposed to varying concentrations of glutamate, erastin, or RSL3 after knockdown with Ct, Nrf2, or Atf4 siRNAs (n = 4/condition). E Levels of NRF2, ATF4, and actin in MC65 cells without (− Aβ) and with (+ Aβ) intracellular Aβ aggregation after 1 day (d1) and 2 days (d2), assessed by western blotting and quantified (n = 3/condition). F Levels of NRF2, ATF4, and actin after knockdown with control (Ct), Nrf2, or Atf4 siRNAs (n = 3/condition). G Survival of MC65 cells exposed to Aβ toxicity after knockdown with Ct, Nrf2, or Atf4 siRNAs 3 days later (n = 4–5/condition). Two-way repeated measures ANOVA and Tukey’s multiple comparisons test, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. All data are mean ± SD