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Figure 5. XBcl9 subcellular localization and levels correlate with dorsal development. A-C: Stage 7, 8.5, and 9.5 embryos bisected into dorsal and ventral halves were immunofluorescently stained for endogenous XBcl9 protein (green) using polyclonal antiserum. A(ii, iv), B(ii, iv), and C(ii, vi) are preimmune serum negative controls for XBcl9 staining. C (iii, iv, vii, and viii) are magnified images of nuclei expressing XBcl9. DAPI (4 prime -6-diamidino-2-phenylindole) staining specifically identifies nuclei. D: Quantitative representation of the fraction of nuclei expressing XBcl9 in 8-12 dorsal (blue) and ventral (red) embryo halves. Data from prestage 8 (stage 7) embryos were not included, because we found no evidence of nuclear staining at this stage. E: XBcl9, X beta -catenin, and XGSK3 beta levels in D2O dorsalized or ultraviolet ventralized embryos. Total protein was standardized to beta -tubulin levels. DAI, dorsoanterior index; n = sample size. Scale bar = 150 mu m in A,B,C (i-ii and v-vi), 200 mu m in C (iii-iv and vii-viii). |
