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. 2010 Dec 3;286(10):7947–7957. doi: 10.1074/jbc.M110.156430

FIGURE 4.

FIGURE 4.

Phosphorylated p38MAPK and activated ATF6α cooperate to enhance the expression level of BiP in dopaminergic neurons treated with MPP+. A–E, immunoblot of cell lysates from SH-SY5Y cells treated with 1 mm MPP+ for 2, 6, 12, 24 h using anti-ATF6α, phosphorylated PERK (p-PERK), PERK, p38MAPK, phosphorylated p38MAPK (p-p38MAPK), BiP, and β-actin antibodies. 5 μm SB203585 was added into the medium prior to MPP+ treatment for the indicated times. Densitometry of ATF6α (N) (B), p-PERK (C), p-p38MAPK (D), and BiP (E) was performed on scanned immunoblots images using the Image-J and normalized to β-actin, PERK, p38MAPK, and β-actin, respectively. Each bar denotes the mean ± S.D. F, BiP promoter-Luciferase plasmid and pRL-SV40 plasmid were transfected into SH-SY5Y cells with p38MAPK overexpression vector or Mock vector. Each group of transfected cells was treated with or without 1 mm MPP+ for 0.5, 1, 2, 4, 12, 24 h. Relative activity is defined as the ratio of firefly luciferase activity to Renilla luciferase activity in dual-luciferase assay. G, dopaminergic neurons from primary co-culture of the midbrain and the striatum of ATF6α WT(+/+) or ATF6α KO(−/−) mice transfected with BiP promoter-Luciferase vector and pRL-SV40 vector and then treated with 1 mm MPP+ for 24 h with or without 5 μm SB203585. Relative activity is defined as the ratio of firefly luciferase activity to Renilla luciferase activity in dual-luciferase assay.