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Supplementary Figure 3: (A) Graph showing percentage of mitotic cells in an arbitrary area on the injected side of embryos injected with Xic1 mRNA constructs as labelled, compared to the uninjected side taken to be 100%. Error bars represent standard deviation, n=5 embryos. Levels of injected mRNAs were kept at a low enough level so the cell cycle was only substantially slowed and not completely inhibited; when levels of Xic1 are high enough to completely block cell cycling, this results in extensive apoptosis and embryonic death (8 and data not shown). The extent of cell cycle inhibition in these embryos was measured by monitoring pH3 expression on the injected versus the uninjected side of stage 15 embryos, by whole-mount antibody staining. The full-length, N-terminus and NT 35-96 Xic1 constructs can all inhibit overall CDK kinase activity8,9 and substantially slow the cell cycle by similar amounts. The C-terminus of Xic1 can also slow the cell cycle, although somewhat less effectively, and it does so by inhibiting the replication factor PCNA. (B) Injection of Xic1 mRNA constructs alone does not alter size of the heart compared to uninjected controls at stage 29/30, as determined by TIc expression, n=28-56 embryos per treatment. Image published in: Movassagh M and Philpott A (2008) Published on behalf of the European Society of Cardiology. 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-NonCommercial license Larger Image Printer Friendly View

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