Fig. 5. Effect of Rab10 on ciliogenesis in SN4741 cells. (A) The cells were transfected with GFP or GFP-Rab10 plasmids and after a day, they were exposed to serum starvation for 24 h. (A) Ciliated cells were stained with Arl13B (red) antibody and Arl13B+ ciliated cells were counted in non-transfected (none) and GFP+ or GFP-Rab10+ transfected cells (green). A total of 320~413 cells in ten images were analyzed. A representative image of double-stained GFP-Rab10 transfected cells is shown. GFP-Rab10 cells transfected and non-transfected with cilia are indicated by an arrow and arrowheads, respectively. The transfection efficiency of GFP and GFP-Rab10 was 32 and 47%, respectively, and the average was 40%. Scale bar: 50 μm. B. Western blot analysis of each transfected cell type. The cell lysates were subjected to Western blot analysis with GFP and Rab10 antibodies. GFP antibody detected both GFP and GFP-Rab10 (red arrow) whereas Rab10 antibody detected both endogenous (endo) Rab10 and exogenous (exo) GFP-Rab10 proteins. *Indicates a non-specific band. n.s.: not significant.
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