HT-22 cells were treated with 5 μM tunic for 2 h followed by washing in PBS then incubated in fresh media for 24 h. After this period, the neuron conditioned media (NCM) was collected. (A) HT-22 cells were cultured in NCM from UT or tunic treated HT-22 cells for 6 h or treated directly with tunic (5 μM) for 6 h followed by measurement of CHOP and GRP78 by qPCR. N = 3 independent cell culture preparations, *P ≤ 0.05, **P ≤ 0.01, determined by one-way ANOVA with Tukey’s multiple comparisons test. (B) Astrocytes were cultured in NCM from UT or tunic treated HT-22 cell for 6 h followed by immunoblotting. (C) Primary microglia were treated with NCM as in B, followed by immunoblotting. Astrocyte lysate (astro, lane 4) was included to confirm efficient separation of microglia and astrocytes.