Table 2.

Effect of Different PTK Inhibitors on Zygote Development.

Treatment	Stage Treated	n	% 2-cell	S.E.M.
DMSO	MII/anaphase	80	82.1	0.045
PD168393 (5μM)	“	49	80.1	0.071
GTP14564 (20μM)	“	55	78.9	0.071
PP2 (10μM)	“	34	72.5	0.185
(100μM)	“	37	33.7*	0.086
SKI-606 (1μM)	“	72	72.3	0.154
(2.5μM)	“	21	43.5*	0.165
(10μM)	“	89	14.9*	0.086
(10μM)	pronuclear	106	13.3*	0.033
c-Fyn RNA (1.5μg/ul)	“	77	83.2	0.650
dn-Fyn RNA (1.5μg/ml)	“	69	37.9*	4.550

Mature, MII oocytes were collected from superovulated females and fertilized by incubation with capacitated sperm for 4.5 hours as described in “Materials and Methods”. The eggs were then transferred to small droplets of mKSOM^AA containing the indicated inhibitor and overlain with oil equilibrated with the same inhibitor. This transfer was done either immediately after fertilization (MII/anaphase) or at the early pronuclear stage (pronuclear). For RNA injection, mRNA was transcribed in vitro and prepared as previously described (Sharma, et al., 2005), then pronuclear stage zygotes were injected with approximately 1–2pg of RNA and cultured in normal mKSOM^AA. The status of the zygotes was assessed at 24 hours by examination by Hoffmann modulation or confocal fluorescence microscopy to establish whether sperm incorporation did occur and whether cell division had occurred. Values represent the mean from at least two experiments (*) indicates that the value is significantly different from the DMSO control group as determined by t test (P<0.05).