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Fig. 2. PtdIns(4,5)P₂ regulates adhesion downstream of PLD activation. (A) Both in the absence or presence of 0.5% butan-1-ol and following treatment with either 100 µM PtdCho, 100 µM PtdOH, 50 µM DAG, 100 µM PtdIns(4,5)P₂ or vehicle (control) the level of cellular adhesion to serum-coated plastic was quantified. Optimal concentrations of each lipid were determined in dose-response experiments. The dotted line represents the level of adhesion observed in control cells not treated with butan-1-ol. In the presence of butan-1-ol, treatment with lipid was considered to have completely recovered the loss of adhesion due to PLD inhibition if the level of adhesion was above this line (B) Adhesion to serum-coated plastic was quantified after cells had been maintained in suspension for the indicated time points and subsequently treated with PtdOH, PtdIns(4,5)P₂ or vehicle (no lipid). Using a Student's t-test, P values of <0.05% were obtained (* and **). The level of adhesion for each individual lipid was obtained from an average of at least three experiments and error bars represent the standard deviation from the mean.

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