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Fig. 3. Expression analysis of x-Bic-C. (A-K) Localization of xBic-C mRNA analyzed by in situ hybridization. (A) Albino oocytes at stages II, III and V. (B) In situ hybridization of a stage V oocyte section, inset shows a magnification of the punctate staining in the oocyte vegetal cortex. Low levels of xBic-C transcripts are found in the animal cytoplasm, but not in the germinal vesicle nucleus. (C) Double in situ hybridization showing vegetal expression of xBic- C and of the germ plasm marker Xcat-2 (Forristall et al., 1995); the arrowhead indicates Xcat-2. Although both genes are localized to the vegetal pole, their distribution is clearly different. (D) Regularly cleaving embryos with a strong dorsoventral polarity at the 4-cell stage. Note the dorsal (light blastomeres) displacement of xBic-C mRNA upon cortical rotation. (E,F) Hemisections of stage 8 and 10.5 embryos on a plane perpendicular to the dorsoventral axis. (G,H) Stage 12 hybridized as a whole mount and as a hemisection; note zygotic expression in the dorsal lip (arrowhead). (I) Stage 24, dorsal view; (J) stage 30, lateral view; staining is seen in the pronephros, the pronephric duct and the posterior spinal cord. (K) Transverse section of a stage 30 embryo showing the staining in the pronephros anlage as well as in the floorplate. (L) RT-PCR analysis of the expression of xBic-C at different developmental stages using ODC as loading control. |
