Additional file 1 of α-Synucleinopathy associated c-Abl activation causes p53-dependent autophagy impairment

Additional file 1: Figure S1. Increased 3-nitrotyrosine (3-NT) immunoreactivity in TgA53T neurons indicate neuronal oxidative stress. Figure S2. Activation of c-Abl and p53 is associated with α-synucleinopathy. Figure S3. Activation of Mdm2 in A53TαSyn Tg Mice brainstem (BST). Figure S4. Increased cytosolic p53 in TgA53T neurons. Figure S5. Colocalization of p62 accumulation with pS129αS in TgA53T. Figure S6. c-Abl activity and autophagy in HEK-293 cells. Figure S7. c-Abl activity modulates nutrient starvation induced autophagy. Figure S8. Constitutively active c-Abl suppresses Mdm2 levels and increases p53 levels. Figure S9. Increased mTOR activation with c-Abl activation. Figure S10. c-Abl-dependent effects on mTOR and AMPK is blocked by inhibition of p53. Figure S11. Inhibition of c-Abl by nilotinib (Nilo) modulate basal autophagy, in neuronal cells. Figure S12. c-Abl-dependent effects on mTOR and AMPK. Figure S13. Nilotinib treatment decreases activation of astrocyte and microglia in the TgA53T Cerebellum and Brainstem. Figure S14. Nilotinib treatment is not able to inhibit c-Abl in the TgA53T spinal cord. Figure S15. Nilotinib treatment is not able to change αS pathology or attenuates autophagy deficit in spinal cord. Figure S16. Nilotinib treatment is not able to inhibit astrocyte/microglia activation in spinal cord. Table S1. Antibodies used for Western blotting and immunohistochemistry.

附加文件1：
图S1：TgA53T神经元中3-硝基酪氨酸（3-nitrotyrosine，3-NT）免疫反应性增强，提示神经元存在氧化应激。
图S2：c-Abl与p53的激活与α-突触核蛋白病（α-synucleinopathy）相关。
图S3：A53TαSyn转基因（Transgenic，Tg）小鼠脑干（Brainstem，BST）内Mdm2的激活。
图S4：TgA53T神经元胞浆内p53水平升高。
图S5：TgA53T模型中p62聚集与pS129αS存在共定位现象。
图S6：HEK-293细胞中的c-Abl活性与自噬（autophagy）情况。
图S7：c-Abl活性可调节营养剥夺诱导的自噬。
图S8：组成型激活的c-Abl可降低Mdm2水平并升高p53水平。
图S9：伴随c-Abl激活的雷帕霉素靶蛋白（mTOR）激活水平升高。
图S10：c-Abl对mTOR与腺苷酸活化蛋白激酶（AMP-activated protein kinase，AMPK）的依赖效应可被p53抑制所阻断。
图S11：尼洛替尼（nilotinib，Nilo）对c-Abl的抑制作用可调节神经元细胞的基础自噬水平。
图S12：c-Abl对mTOR与AMPK的依赖效应。
图S13：尼洛替尼处理可降低TgA53T小鼠小脑与脑干内星形胶质细胞与小胶质细胞的激活程度。
图S14：尼洛替尼处理无法在TgA53T小鼠脊髓中抑制c-Abl活性。
图S15：尼洛替尼处理无法改变αS病理表现，也不能缓解脊髓内的自噬功能缺陷。
图S16：尼洛替尼处理无法在脊髓中抑制星形胶质细胞/小胶质细胞的激活。
表格S1：用于蛋白质印迹与免疫组织化学实验的抗体列表。