Fig. 2

C-terminus of Best1 is important for binding with 14–3-3γ. a Schematic diagram of full-length (Best1-WT), the first C-terminal region (292–380 amino acids, Best1-C1), the second C-terminal region (381–470 amino acids, Best1-C2), the third C-terminal region (471–551 amino acids, Best1-C3), C1 deleted mutant of Best1 (Best1-ΔC1) and S358A mutant of Best1 (Best1-S358A). b Co-IP of HA-14-3-3γ with GFP-Best1-C1 (C1), GFP-Best1-C2 (C2) and GFP-Best1-C3 (C3). Whole lysate of COS7 cells was immunoprecipitated with anti-HA antibody and then analyzed by Western blot with anti-GFP antibody. 5% of the total lysate was used as input for the immunoprecipitation. c Co-IP of HA-14-3-3γ with GFP-Best1-WT (WT) and GFP-tagged S358A mutant of Best1 (S358A). Whole lysate of COS7 cells was immunoprecipitated with anti-HA antibody and then analyzed by Western blot with anti-GFP antibody. 5% of the total lysate was used as input for the immunoprecipitation. d BiFC experiment with VN-14-3-3γ and Best1-VC, where N- and C-terminal halves of Venus fluorescent protein were fused to 14–3-3γ and Best1, respectively. BiFC fluorescent signals were detected by the yellow color. The fluorescent signal was detected at the plasma membrane of the cells by the yellow color. Cells only expressing both Best1-WT-VC and VN-14-3-3γ showed intense fluorescent signal (lower left panel). Scale bar, 10 μm