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The localization of both proteins was unchanged in Pten mutant nerves when immunohistochemically stained in teased fibre preparations (Fig 4C).
We analysed levels of ubiquitinated proteins in P90 Floxed/pltd/plt, P0-Cre nerve lysates and found an increase of polyubiquitinerve proteins in mutant nerves when compared with controlysates 8E).
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This difference is reflected at a later time-point (6 months), when the regenerated distal regions of mutant nerves were visibly smaller.
Instead, peripheral nerve development appeared largely normal in the absence of Nf1 except for abnormal Remak bundles, the nonmyelinated axon-Schwann cell unit, identified in postnatal mutant nerves.
In contrast, Jagged1 was upregulated in Dicer mutant nerves.
In Dicer mutant nerves, we observed a significantly lower Ras expression compared to control nerves.
Next, we performed a time course analysis of changes in Dicer mutant nerves (Fig. 3).
We did not observe normal Remak bundle formation in Dicer mutant nerves.
In contrast, at p18 we observed increased Akt phosphorylation in Dicer mutant nerves.
In parallel to increased proliferation, Dicer mutant nerves at p22 showed increased cell death as determined by TUNEL staining (<0.05% TUNEL positive nuclei in all control nerves and ca. 2% in Dicer mutant nerves).
Furthermore, we observed an increased phosphorylation of Erk in Dicer mutant nerves at p20.
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