Figure 5
Protection of the primary hippocampal neurons by CRT or by treatment with Ca2+/ER stress-modulating compounds. (A,B) WT and Atf6b−/− hippocampal neurons were infected with a control or CRT-expressing lentiviral vector, and the expression levels of CRT and calnexin were measured by western blotting (A). n = 3 experiments. Data are shown as mean ± SEM. *p < 0.05, **p < 0.01 by a two-way ANOVA followed by the Bonferroni tests. Cells were then treated with Tm (1 μg/ml) for 24 h, and cell death was evaluated by immunocytochemical staining for cleaved caspase-3 (B). Scale bar: 20 μm. n = 3 experiments. Data are shown as mean ± SEM. *p < 0.05, ***p < 0.001 by a two-way ANOVA followed by the Bonferroni test. (C) WT and Calr+/− primary hippocampal neurons were treated with Tm (1 µg/ml) for 24 h, and cell death was evaluated by immunocytochemical staining for cleaved caspase-3. n = 3 experiments. Scale bar: 20 μm. Data are shown as mean ± SEM. **p < 0.01 by a two-way ANOVA followed by the Bonferroni tests. (D) WT and Atf6b−/− hippocampal neurons were treated with Tm (1 μg/ml) together with BAPTA-AM (5 µM), 2-APB (2 μM) or salubrinal (5 µM). Cell death was evaluated by immunocytochemical staining for cleaved caspase-3. n = 3 experiments. Typical images are shown in Fig. S5. Data are shown as mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001 by a two-way ANOVA followed by the Bonferroni test.
