Figure 4

Endoplasmic reticulum stress and mesencephalic astrocyte-derived neurotrophic factor expression in cultured primary glial cells. (A to D) Glial cells in the mixed culture were treated as indicated (serum+ control, DMEM medium containing 5% serum; Tm, tunicamycin, 1 μg/ml; MG132, 10 μM; serum–, serum-free DMEM medium). Twenty-four hours after treatment, the cells were collected and processed for RT-PCR and immunoblotting (IB). Levels of mRNAs (A) and proteins (C) were quantitated and normalized by actin. The quantitative data in (A) and (C) are shown in (B) and (D), respectively. Values expressed as mean ± standard error of the mean of three independent experiments. *P <0.05, **P <0.01, and ***P <0.001, compared with serum+ control. (E to P) Glial cells were treated with serum+ (E to G), Tm (H to J), MG132 (K to M), and serum– (N to P). Twenty-four hours after treatment, the cells were stained with 4^′,6-diamidino-2-phenylindole (F, I, L, and O) and propidium iodide (G, J, M, and P), and then observed with a microscope under bright field (E, H, K, and N) and fluorescence. Scale bar = 50 μm.