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. 2010 Oct 5;1(4):e00176-10. doi: 10.1128/mBio.00176-10

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Copyright © 2010 De Miranda et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution-Noncommercial-Share Alike 3.0 Unported License, which permits unrestricted noncommercial use, distribution, and reproduction in any medium, provided the original author and source are credited.

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FIG 2 — Poly(I ⋅ C) treatment during gestation induces defective proliferation in fetal cerebral cortical cells. Pregnant mice were treated with either PBS or poly(I ⋅ C) on 3 consecutive days (GD15 to GD17) and injected with bromodeoxyuridine (BrdU) on GD16. Cerebral cortical cells were isolated from embryos on GD18, and quantitation of BrdU-labeled cells was performed by flow cytometric analysis. (a) Viable cerebral cortical cells were gated according to forward scatter (FSC) and side scatter (SSC) profiles. The percentage of live cells (85.5%) is shown. 50K, 50,000. (b) Representative histogram showing intracellular BrdU incorporation in cerebral cortical cells. The numbers above the brackets indicate the percentages of brightly labeled BrdU cells (BrdU^high) and a subpopulation of cells that incorporated low levels of BrdU (BrdU^low). Gating for the BrdU-positive cells was based on the anti-BrdU antibody staining observed with cells obtained from mice that were not injected with BrdU (negative control). Anti-BrdU antibody conjugated to allophycocyanin (APC) (BrdU-APC) was used. (c) Representative histogram showing the mean fluorescence intensity (MFI) of BrdU-labeled cells represented as a percentage of maximum BrdU incorporation. Values for the different groups are indicated as follows: PBS-treated mice (red line), poly(I ⋅ C)-treated mice (blue line), and negative-control mice (gray line). The experiments were performed with cerebral cortical cells isolated from embryos (n = 7); data from one representative cortex are shown. (d) Percentage of BrdU^high cells in cerebral cortical cells from mice treated with PBS or poly(I ⋅ C). Results are expressed as percentages of total gated cells. The values for the poly(I ⋅ C)-treated group are significantly different (P < 0.05 by the Mann-Whitney U test) compared to the values for the PBS-treated control group as indicated by the bracket and asterisk. The height of the box plot shows the interquartile range, and the horizontal line indicates the median. The range is indicated by the error bars, and the circles represent outlier values.