Figure 4

KIF20A is essential for Golgi scattering induced by SUN1 depletion. (a) SUN1- and SUN2-containing protein complexes were precipitated using anti-SUN1 or anti-SUN2 pAb, resolved by SDS-PAGE, and subjected to silver staining; the bands indicated by arrows, an arrowhead, and an asterisk were analyzed by LC–MS/MS. The bands indicated by arrows with molecular weights of approximately 75 kDa are two SUN2 splicing variants, RefSeq NM_001199580 and RefSeq NM_001199579. (b) Immunoprecipitation was performed using anti-SUN1 (lane 1) or anti-SUN2 pAb (lane 2) and analyzed by Western blotting using anti-SUN2 pAb. (c) Immunoprecipitation was performed using normal rabbit IgG (lane 1) or anti-SUN2 pAb (lane 2) and analyzed by Western blotting using anti-KIF20A pAb. (d) and (e) Cells were transfected with siSUN1 and siRNA against KIF20A (siKIF20A). The cells were then stained with anti-SUN1 pAb and anti-GM130 mAb. Bar, 10 µm. (e) Box-and-whiskers plots represent GM130 labeled Golgi complex area. ** P < 0.01. (f) and (g) Cells were transfected with siSUN1 and incubated for 36 h. Paprotrain was then added. After 12 h, the cells were fixed and stained with anti-SUN1 pAb and anti-GM130 mAb. Bar, 10 µm. (g) Box-and-whiskers plots represent GM130 labeled Golgi complex area. * P < 0.05.