PLD2 antisense oligonucleotides down-regulate PLD2 activity, and acetylcholine and choline levels in SN56 cells. SN56 cells were transfected with either control or antisense oligonucleotides as described in the Experimental Procedures. After 48 hours the cells were preincubated for 20 minutes in choline-free medium and then incubated in fresh choline-free medium containing 50 μM neostigmine with or without 100 nM PMA for 45 minutes. PLD activity in the intact cells (A) and the intracellular choline (B) and acetylcholine (C) content were measured. The data were analyzed by two-way ANOVA and Fisher's post hoc test and are expressed as average ± SEM. The asterisks indicate a significant difference between the antisense and control transfectants (A-C). A. Effect of antisense oligonucleotides on PLD activity. The effect of PMA treatment is statistically significant in all groups. B. Effect of antisense oligonucleotides on intracellular choline level. C. Effect of antisense oligonucleotides on intracellular ACh level. The effect of PMA treatment is statistically significant in all groups.