Figure 1.
Pten expression in MPNST and neurofibroma.
a) Proportion of Pten-positive tumour cells as determined by immunohistochemistry. Each dot represents one tumour. Sporadic MPNST are depicted in grey b) MPNST 29250 with a transition to pNF. Note stronger Pten expression in the pNF (lower part). c) Higher magnification of pNF. Original magnification 200x and 400x. d) Western blot of MPNST and neurofibromas. VEGF concentrations in tumour lysates are indicated.
Figure 2.
Pten expression in nerve tissue (a-c) and Schwann cell cultures (d).
a) Standard DAB staining. b) Double staining for Pten (green) and neurofilament (red). c) Double staining for Pten (green) and S100 (red). Original magnification 400x.
Figure 3.
Western blot analysis of Pten and p-p70S6 kinase (isoform p-p70 of the S6 kinase, indicates mTOR activation) and examination of miR-21 expression by real time PCR a) Pten analysis of MPNST cell lines, dermal fibroblasts and neurofibroma derived Schwann cell cultures (SC1-4).
The grey line indicates mean Pten expression of 4 Schwann cell cultures. b) Quantification of Pten expression (normalized with β-actin). c) Expression of Pten and p-p70S6 kinase in NF1 positive (+/+) and NF1 negative (−/−) Schwann cells. d) Relative expression of miR-21 as determined by real time PCR. * Endogeneous control RNU44 was not detectable. This cell line was thus omitted from analysis. SC = neurofibroma derived Schwann cells.
Figure 4.
Effect of rapamycin on MPNST cell lines and fibroblasts.
a) Dose dependent inhibition of proliferation after 4 days of treatment. b) Pten protein levels correlate with rapamycin sensitivity.