Fig. 4.

Suppression of PLD1 expression in T24 and Calu-1 cells results in apoptosis upon serum withdrawal. (A) T24 and Calu-1 cells were plated at a density of 10⁵ cells/60 mm plate. 24 hr later the cells were shifted to media containing 0.5% serum. 18 hr later the cells were either untreated (Con), or treated with either tertiary BtOH (t-BtOH) or primary BtOH (1-BtOH) and cell viability (upper graph) and PARP cleavage (lower blot, Cl PARP) was determined 6 hr later by Western blot analysis. Error bars represent the standard deviation for duplicate samples from a representative experiment repeated at least two times.

(B) T24 and Calu-1 cells were plated at a density of 10⁵ cells/60 mm plate. 24 hr later the cells were transfected with either control (Con) scrambled siRNA or siRNA specific for PLD1 as in Fig. 2. 24 hr later the cells were shifted to media containing 0.5% serum. 18 hr later cell viability (upper graph) was determined as in Fig. 4. In the lower panel, PLD1 levels and PARP cleavage (Cl PARP) were determined using Western blot analysis. Error bars represent the standard deviation for duplicate samples from a representative experiment repeated at least two times.