BACKGROUND Flotillin-2 (Flot2) is a lipid raft scaffold protein that is thought to be related to neural differentiation. Flot2 is phosphorylated by Fyn, a Src kinase, and causes raft-dependent endocytosis; however, the exact role of Flot2 in neural differentiation remains unclear. To reveal the roles of lipid raft-associated proteins during neural differentiation, we tried to analyze the expression and localization. RESULTS In this study, we found that the expression levels of the Flot2 and Fyn proteins increased in whole-cell lysates of P19C6 cells after neural differentiation. In addition, sucrose density fractionation and immunofluorescence experiments revealed an increase in the localization of Flot2 and Fyn to lipid rafts after neural differentiation. We also found that Fyn partially colocalized with Flot2 lipid rafts in neural cells. CONCLUSION The observed distribution of Fyn and level of inactivated Fyn and/or c-Src in detergent-resistant membrane (DRM) fractions suggests that the amount of activated Fyn might increase in DRM fractions after neural differentiation. Overall these findings suggest that Flot2 lipid rafts are associated with Fyn, and that Fyn phosphorylates Flot2 during neural differentiation of P19C6 cells.

中文翻译：

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脂质筏的Flot2组件在P19C6细胞神经分化过程中改变了定位。

背景技术Flotillin-2（Flot2）是一种脂筏支架蛋白，被认为与神经分化有关。Flot2被Src激酶Fyn磷酸化，并引起筏依赖性细胞吞噬作用。然而，Flot2在神经分化中的确切作用仍不清楚。为了揭示脂质筏相关蛋白在神经分化过程中的作用，我们试图分析其表达和定位。结果在本研究中，我们发现神经分化后P19C6细胞全细胞裂解物中Flot2和Fyn蛋白的表达水平增加。此外，蔗糖密度分级和免疫荧光实验显示神经分化后Flot2和Fyn在脂筏上的定位增加。我们还发现Fyn与Flot2脂质筏在神经细胞中部分共定位。结论观察到的Fyn分布以及去污剂抗性膜（DRM）组分中失活的Fyn和/或c-Src的水平表明，神经分化后，DRM组分中活化的Fyn的量可能增加。总体而言，这些发现表明Flot2脂质筏与Fyn相关，并且Fyn在P19C6细胞的神经分化过程中使Flot2磷酸化。