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Figure 5. Bcl-xL regulation of NF-κB RNAs:A: RNA was extracted from eggs and embryos at various stages and analyzed by RT-PCR (28 cycles); levels of RelA, Rel2, Rel3, RelB and Xp100 RNAs drop between stage 7 and 9 and, except for RelB, increase following gastrulation (stage 12/13). Levels of Bcl-xL RNA appear relatively constant throughout this period of development. B: At stage 16, embryos were dissected into anterior dorsal (AD), posterior dorsal (PD), anterior ventral (AV), and posterior ventral (PV) quadrants and RNA was analyzed by RT-PCR; RelB was not expressed at this stage; expression of RelA, Slug and Sox9 are restricted to the anterior dorsal quadrant, while Bcl-xL, Rel2, Rel3, and Xp100 RNAs can be detected throughout the embryo. C, D: Animal caps were prepared from embryos injected with GR-Bcl-xL-GFP RNA (“GRBclxL”)(600 pg/embryo) and either left untreated (0.1%DMSO)(“Con”), treated with 20 µM dexamethasone (“+Dex”), treated first with 100 µg/ml emetine and then dexamethasone (“+Dex+Eme”), or treated with emetine alone (“+Eme”), and analyzed at stage 11 for Slug, RelA (C), Rel2 and Rel3 (D) RNA levels. Treatment with emetine blocked the increase in Slug and Rel2, but not RelA and Rel3 RNAs; emetine treatment alone produced control or slightly reduced levels of Slug and Rel RNAs. E: Activation of the GR-Bcl-xL-GFP protein in embryos injected with the Slug MO produces an increase in the level of Snail RNA, analyzed at stage 11. F: In animal caps derived from GR-BclxL-GFP RNA injected embryos, AKBA (50 µM) inhibited the dexamethasone-induced increases in Slug, Snail, and RelA RNA levels; while treatment with AKBA alone lead to a decrease in Slug, Snail and RelA RNA levels. G: In animal caps, injection of RelAΔSP RNA (600 pg/embryo) blocked Rel3 and Xp52 RNA induced activation of the 3XκB reporter. H: The ability of Bcl-xL RNA to increase levels of RelA and Slug RNAs in animal caps was blocked by the co-injection of RelAΔSP RNA. Error bars in C–H reflect standard deviation from the mean of multiple experiments. Image published in: Zhang C et al. (2006) Zhang et al. This image is reproduced with permission of the journal and the copyright holder. This is an open-access article distributed under the terms of the Creative Commons Attribution license Larger Image Printer Friendly View

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