Fig. 1. Ciliogenesis in dSH-SY5Y cells and rat cortical neurons after MLi-2 treatment. Ciliated cells and nuclei were stained with Arl13B antibody and Hoechst 33342, respectively. Arl13B+ (green) cells were counted as ciliated cells and divided by the total number of cells to obtain the percentage of ciliated cells. In every cell image, the nuclei were stained with Hoechst 33342 (blue). (A) dSH-SY5Y cells incubated with DMSO (-) or 100 nM MLi-2 under serum starvation (S.S) or normal (Ctrl) conditions for 24 h. Scale bar: 20 μm. A total of 393~687 cells in nine images were analyzed. A summary graph and each representative image are shown. #p<0.05 by the t-test. Primary rat cortical (B) and hippocampal (C) neurons treated with DMSO (-), 1 μM GSK-2578215A (GSK, B), or 100 nM MLi-2 (C) for 24 h. The treatment scheme, summary graph, and a representative image for each cell type are shown. Scale bar: 100 μm. A total of 81~92 cells in eleven images were analyzed. The white arrow indicates a ciliated cell. (D) The decrease in LRRK2 kinase activity in hippocampal neurons treated with MLi-2 was confirmed by a decrease in pRab10 levels. n.s.: not significant.
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