PMC

Normal nervous tissue and brain served as control. PCR was performed in triplicates and accepted when Ct value variation was less than 0.5 cycles. MPNST with gene amplification are marked with *.

A) Amplification pattern of 9 genes in an MPNST and its corresponding cell line S462. Sunitinib targets are highlighted in grey. B) Transcript and protein levels in 5 MPNST cell lines, dermal fibroblasts and neurofibroma derived Schwann cells (SC).

A) Dose dependent inhibition of MPNST cell lines after 4 days of treatment. B) For better comparability of cell lines treatment with 1.0 µM sunitinib is depicted in the bar chart. The dashed line marks IC₅₀. C) Detection of apoptosis in S462 cells treated for 3 days with 10µM sunitinib. Note formation of apoptotic bodies by DAPI staining (upper panel) and DNA fragmentation as detected by the Apoptag assay (lower panel).

A) Transcript levels of KDR (VEGFR-2) and VEGF. Corresponding concentrations of VEGF in cell lysates is shown below the bar chart (nt: not tested; ND: not detected). On the right side cytochemistry of VEGF and VEGFR-2 in S462 cells is shown. B) Serum starved MPNST cell lines ST88-14 and S462 were stimulated with PDGF-AA (50ng/ml). Where indicated cells were pre-incubated with 5µM sunitinib for 1h to block signaling. C) Serum starved HUVECs were stimulated with VEGF-165 (100ng/ml) or HUVEC medium (HM). D) VEGF-165 (100ng/ml) was used to stimulate serum starved NSF1 cells. Sunitinib blocked VEGF induced signaling. E) VEGF-165 and conditioned S462 medium (CM) was used to stimulate serum starved S462 cells. EGF (100ng/ml) served as positive control. Cells were pre-incubated with 5µM sunitinib where indicated. Densidometric analysis of pMAPK/total MAPK is shown in the bar chart.