TABLE 2.

Inhibition of VEGF-stimulated DNA synthesis in T24 cells by a DN-SPK mutant (SPK1-G81D)^a

Agent	% of BrdU-positive cells among SGFP-positive cells		% of BrdU-positive cells among SGFP-negative cells
	Without VEGF	With VEGF	Without VEGF	With VEGF
Empty vector	17	82	19	82
Wt-SPK	21	80	23	74
SPK-G81D	16	44	27	77

^aT24 cells were cotransfected with pSGFP and pCMV5/myc-1, pCMV5/myc-1-mSPK1(Wt), or pCMV5/myc-1-mSPK1-G81D. VEGF stimulation of DNA synthesis was determined by assaying BrdU incorporation by immunofluorescence microscopy. The percentages of cells exhibiting BrdU incorporation (DNA synthesis) in transfected cells (SGFP positive) and untransfected cells (SGFP negative) were determined with 100 cells under each condition. Relative to cells transfected with the vector control or the plasmid harboring Wt-SPK, the cells transfected with SPK1-G81D exhibited approximately 50% inhibition of VEGF-induced DNA synthesis (P < 0.006). Relative to SGFP-negative cells in the same dish, SGFP-positive cells cotransfected with DN-SPK exhibited a 42% inhibition of VEGF-induced DNA synthesis (P < 0.004).