Figure 3. Aβ1–42‐dependent Atf4 localization requires immediate axonal signaling.
- Hippocampal neurons were cultured in microfluidic chambers for 10–11 DIV, and axons were transfected with scrambled or Atf4‐targeting siRNA. 24 h after transfection, axons were treated with vehicle or Aβ1–42 for 6 h. Axons were immunostained for ATF4 and βIII‐tubulin. Mean ± SEM of 30 optical fields per condition (n = 6 independently performed experiments per group). *P < 0.05; ns, not significant; Holm–Sidak multiple t‐tests.
- Hippocampal neurons were cultured in microfluidic chambers for 11–12 DIV and axons were treated with vehicle, emetine (100 nM), or ciliobrevin A (5 μM) for 45 min. Axons were then treated with vehicle or Aβ1–42 for 6 h. Neurons were fixed and Atf4 transcript levels were measured using quantitative FISH. The background fluorescence was determined using a non‐targeting Gfp probe set (neg. probe) and set to zero. Mean ± SEM of 26–30 optical fields per condition (n = 6 independently performed experiments per group). *P < 0.05; ns, not significant; Holm–Sidak multiple t‐tests.