Figure 5.

Interaction of the C terminus of AF-6 and profilin. (Top) Two-hybrid interaction. An isolated profilin I cDNA containing an additional leader sequence and PCR-amplified profilin I and profilin II cDNAs fused to GAD were cotransformed with LBD AF-6N into L40 and were assessed for β-galactosidase expression and for their ability to grow on medium lacking histidine. pLex-Lamin and pGADGH were used as negative controls. (Middle) Structure of AF-6. (Bottom) AF-6–profilin association examined in GST pull-down assays. Overlapping fragments of the AF-6 C-terminal 700 amino acids were expressed as GST fusions and used to bind profilin from a whole-cell MDCK lysate. Bound profilin was visualized with an α-profilin Ab in a subsequent Western blot analysis.