Fig. 11. Biochemical confirmation of KIF13B inhibition of ARF6 GAP activity of centaurin-₁. (A) COS cells were transfected with the indicated expression plasmids. 2 days later cells were serum starved, incubated with or without EGF (200 ng/ml). The cells were then lysed and ARF6_GTP precipitated using the GST-MT2A coupled to glutathione beads. The precipitates were then immunoblotted (IB) with an anti-HA antibody. The cell lysates (input) were also immunoblotted using anti-HA antibody, anti-GFP and anti-FLAG antibodies to determine HA-ARF6, FLAG-centaurin₁ and GFP-KIF13B levels respectively. (B) Quantification of data obtained from three similar experiments as shown in A above. Data are the means±s.e.