Table 2.

Distribution of the polar plasm components in the embryos injected with the anti-mtlrRNA ribozymes

RNAs and proteins detected*	Rbzs†	Total no. of embryos scored	No. of embryos without signal in polar plasm, %	Significance‡
gcl	+	324§	6  (1.9)	P > 0.2
	−	320§	9  (2.8)
osk	+	296¶	0  (0)	P > 0.2
	−	239¶	1  (0.4)
VAS	+	1,042‖	73  (7.0)	P > 0.2
	−	944‖	66  (7.0)
TUD	+	300**	13  (4.2)	P > 0.2
	−	313**	18  (5.8)

^*Whole-mount in situ hybridization of the ribozyme-injected embryos using a double-stranded DNA probe for mtlrRNA, osk and gcl mRNA was performed. Immunostaining for VAS and TUD protein was carried out. 

^†Embryos were injected with the anti-mtlrRNA ribozymes (+) or DW (−). 

^‡Probability was calculated vs DW-injected embryos by Fisher’s exact probability test. 

^§The injected embryos were stained with gcl probe. To exclude embryos in which polar plasm leaked out or was delocalized by the injection procedure, the injected embryos were stained with an antibody against VAS protein. The number of embryos without gcl signal was counted among the embryos that showed normal VAS staining. 

^¶ The injected embryos were stained with osk probe. As an internal control for in situ hybridization, the embryos also were hybridized with a DIG-labeled probe detecting bcd mRNA that localizes at the anterior pole region of early embryos. And the injected embryos also were immunostained with an anti-VAS antibody. The number of embryos without osk signal was counted among the embryos that showed normal bcd and VAS staining. 

^‖The injected embryos were immunostained with an anti-VAS antibody. The number of embryos without VAS signal was counted among the injected embryos. 

^**The injected embryos were immunostained with an anti-TUD antibody. The number of embryos without TUD signal was counted among the injected embryos.