Data for: Seipin deletion in mice enhances phosphorylation and aggregation of tau protein through reduced neuronal PPARγ and insulin resistance

We attached the independent replicates of Western blot analyses in the supplementary section that were used in various quantifications. The regions of line rectangles in these blots/gels, as representative images, were used in Figures of the main Text. In the Fig. 1e: the levels of hippocampal NF-H protein in seipin-sKO mice (sKO) and seipin-nKO mice (nKO) were lower than those of WT mice. In the Fig. 2a: the levels of tau protein in hippocampus of seipin-sKO mice, seipin-nKO mice and seipin-aKO mice (aKO) were not altered. In the Fig. 2b: the levels of oligomer tau protein were increased in seipin-sKO mice and seipin-nKO mice. In the Fig. 2c: the levels of hippocampal tau phosphorylation at Thr212/Ser214 and Ser202/Thr205 were increased in seipin-sKO mice and seipin-nKO mic. In the Fig. 3a: the levels of PPARγ protein were decreased in hippocampus of seipin-sKO mice and seipin-nKO mice. In the Fig. 3b: the administration of PPARγ agonist rosiglitazone (rosi) for 7 days could correct the increased tau phosphorylation at Thr212/Ser214 and Ser202/Thr205 in seipin-sKO mice and seipin-nKO mice. In the Fig. 4a: seipin-sKO mice and seipin-nKO mice showed an increase in the GSK3β phosphorylation at Tyr21 and a decrease at Ser9, which were corrected by rosi. In the Fig. 4b: the treatment with AR-A014418 (AR) corrected the increased tau phosphorylation at Thr212/Ser214 and Ser202/Thr205 in seipin-nKO mice. In the Fig. 5a&5b: the phosphorylation of Akt and mTOR was elevated in seipin-sKO mice and seipin-nKO mice, which were corrected by rosi. In the Fig. 5c&5d: seipin-sKO mice and seipin-nKO mice showed a decrease in the ratio of LC3II/I and an elevation of p62 protein, which was rescued by the PI3K inhibitor LY294002 (LY) and mTOR inhibitor rapamycin (Rap), but not AR. In the Fig. 5e&5f: the administration of LY and Rap reduced the levels of oligomer tau protein and tau phosphorylation at Thr212/Ser214 and Ser202/Thr205 in seipin-nKO mice. In the Fig. 6c: the increased phosphorylation of JNK in seipin-sKO mice were corrected by treatment with rosi for 30 days. In the Fig. 6d: the P38 phosphorylation in seipin-sKO mice was not altered. In the Fig. 6e: the increased phosphorylation of tau at Ser396 in seipin-sKO mice was prevented by treatment with rosi for 30 days and the JNK inhibitor SP600125 (SP).

本研究将用于各类定量分析的独立重复蛋白质印迹（Western blot）实验结果附于补充材料中。上述印迹/凝胶电泳图中的矩形框选区域作为代表性图像，被用于正文章节的各图版。
在图1e中，seipin特异性敲除小鼠（seipin-sKO）与seipin神经元特异性敲除小鼠（seipin-nKO）的海马组织中神经丝蛋白H亚基（NF-H）水平均低于野生型（Wild Type, WT）小鼠。
在图2a中，seipin-sKO、seipin-nKO及seipin-aKO小鼠（aKO）的海马tau蛋白水平未发生显著变化。
在图2b中，seipin-sKO与seipin-nKO小鼠的tau寡聚体蛋白水平显著升高。
在图2c中，seipin-sKO与seipin-nKO小鼠的海马tau蛋白在Thr212/Ser214及Ser202/Thr205位点的磷酸化水平显著升高。
在图3a中，seipin-sKO与seipin-nKO小鼠的海马组织中过氧化物酶体增殖物激活受体γ（PPARγ）蛋白水平显著降低。
在图3b中，给予seipin-sKO与seipin-nKO小鼠PPARγ激动剂罗格列酮（rosiglitazone, rosi）干预7天，可逆转其海马tau蛋白在Thr212/Ser214及Ser202/Thr205位点的磷酸化水平升高表型。
在图4a中，seipin-sKO与seipin-nKO小鼠的糖原合成激酶3β（GSK3β）在Tyr21位点的磷酸化水平升高，而在Ser9位点的磷酸化水平降低，上述异常均可被罗格列酮（rosi）逆转。
在图4b中，使用AR-A014418（AR）干预可逆转seipin-nKO小鼠海马tau蛋白在Thr212/Ser214及Ser202/Thr205位点的磷酸化水平升高。
在图5a与5b中，seipin-sKO与seipin-nKO小鼠的蛋白激酶B（Akt）与哺乳动物雷帕霉素靶蛋白（mTOR）磷酸化水平均升高，该异常可被罗格列酮（rosi）逆转。
在图5c与5d中，seipin-sKO与seipin-nKO小鼠的LC3II/I比值降低，p62蛋白水平升高；使用磷脂酰肌醇3-激酶（PI3K）抑制剂LY294002（LY）与mTOR抑制剂雷帕霉素（rapamycin, Rap）可挽救上述异常，但AR无此效果。
在图5e与5f中，给予seipin-nKO小鼠LY与Rap干预，可降低其tau寡聚体蛋白水平以及tau蛋白在Thr212/Ser214、Ser202/Thr205位点的磷酸化水平。
在图6c中，seipin-sKO小鼠的c-Jun氨基末端激酶（JNK）磷酸化水平升高，经罗格列酮（rosi）干预30天后该异常可被逆转。
在图6d中，seipin-sKO小鼠的P38丝裂原活化蛋白激酶磷酸化水平未发生显著变化。
在图6e中，seipin-sKO小鼠的tau蛋白在Ser396位点的磷酸化水平升高，经罗格列酮（rosi）干预30天以及使用JNK抑制剂SP600125（SP）处理后，该异常可被阻断。