Effect of Kif2A-GFP overexpression on AGAP1 knockdown and effect of FLAG-AGAP1 overexpression on Kif2A knockdown. Spreading of HeLa cells treated with siRNA targeting the noncoding region of AGAP1 or Kif2A and transfected with plasmids for expression of either Kif2A-GFP, Kif2C-GFP, FLAG-AGAP1, or ArfGAP1-HA was determined, and data analysis was carried out as in Fig. 7. A, effect of AGAP1 overexpression on Kif2A knockdown. Control Si + empty vector versus Kif2A Si + empty vector, p < 0.001; Kif2A Si + empty vector versus Kif2A Si + FLAG-AGAP1, p < 0.001; Kif2A Si + empty vector + ArfGAP1-HA, not significant. B, effect of Kif2A-GFP or Kif2C-GFP overexpression of AGAP1 knockdown. Control Si + empty vector versus AGAP1 Si + empty vector, p < 0.001; AGAP1 Si + empty vector versus AGAP1 Si + Kif2A-GFP or AGAP1 Si + Kif2C-GFP, not significant. C, effect of AGAP1 and AGAP1 mutants on Kif2A knockdown. The results were analyzed by ANOVA with post-test multiple comparisons. Control Si + empty vector versus Kif2ASi + empty vector, p < 0.0001; control Si + empty vector versus Kif2ASi + FLAG-AGAP1, not significant; Kif2A Si +empty vector versus Kif2A Si + FLAG-AGAP1, p < 0.0001; Kif2A Si +empty vector versus Kif2A Si + FLAG-GLDPH, not significant; Kif2A Si +empty vector versus Kif2A Si + FLAG-ZA, not significant; Kif2A Si +empty vector versus Kif2A Si + FLAG-[R599K]AGAP, p < 0.01; Kif2A Si +FLAG-AGAP1 versus Kif2A Si + FLAG-[R599K]AGAP, p < 0.05. Results shown are the mean ± S.E. from three independent experiments.