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Figure 1. Geminin is Required for Zygotic Gene Expression. (A) Immunoblot showing complete depletion of Geminin by anti-Geminin morpholino oligonucleotides (α-Gem MOs). XRBs, cross-reacting bands serving as a loading control. Geminin runs as a tightly spaced doublet at ~35 kD. (B) Real-time PCR (RT-PCR) data showing decreased expression of most genes in Geminin-depleted embryos at stage 10.5. The expression level if each RNA was normalized to the expression in control embryos (Co). n, number of independent experiments; asterisk indicates P<0.001 (paired t-test); NS, not significant. (C) In situ hybridization for Gsc (C) and Xbra (F) at stage 10.5 showing that depleting Geminin preserves the pattern of expression for both genes. Both cells of two-cell Xenopus embryos were left uninjected (Co) or injected with either anti-Gem MOs or α-amanitin. (I) Xbra expression is affected only in Geminin-depleted cells. One cell of a two-cell embryo was injected with anti-Geminin MOs (arrowhead) and in situ hybridization for Xbra was performed when the embryo reached stage 10.5. Note the markedly larger cells on the injected side (dotted line), indicating that the cells have stopped dividing. |
