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When touched by microtubules, the female pro-nucleus was rapidly pulled towards the male pro-nucleus.
After sperm entry, the male pro-nucleus was pushed by microtubules towards the center of the egg (Fig. 2; Video S3).
Transgenic mice were generated by pro-nucleus injection (FVB/N background).
These constructs were microinjected into the pro-nucleus of the fertilized eggs from CD-1 as described (Nagy et al., 2003) Genotyping of CAG-Dkk1 transgenic mice obtained using the PCR primers CAG-pro (5′-TAGAGCCTCTGCTAACCATGTTCATGCCTT-3′) and CAG-Dkk1 (5′-TCAGCGCAAGGGTAGGGCTGGTAGTTGTCA-3′), yielding 443 bp.
Heterokaryons were visualized by DNA staining of the nucleus (To-Pro-3 iodide) and/or α-tubulin staining.
The total number of cells (neuronal and non-neuronal) in striatal sections from C57Bl/6J WT mice was assessed by counting TO-PRO-3 labeled nuclei.
A total of 45 images of TO-PRO-3 stained nuclei from striatal neurons (9 for each neuronal type) were analyzed.
Next, the percentage of neurons in each nuclear staining category (A E or X, as described above), was calculated with respect to the total number of TO-PRO-3 labeled nuclei.
We then counted for each double staining the number of cells identified by their TO-PRO-3-stained nuclei that were NeuN-positive or DARPP-32-positive (Fig. 4C).
The samples were then analyzed by obtaining z-stacks of TO-PRO-3-stained nuclei using confocal microscopy.
Xie et al. elegantly demonstrate that this phenotype is due to the ectopic expression of Prospero (Pros) in the nuclei of newly generated INPs.
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