Additional file 2. Figure S1 (A) immunoblot for neurofibromin protein in the MPNST cell lines used in this study. Blots were probed with an anti-actinin antibody to compare loading. (B) NF1 mutational status of the MPNST cell lines. (C) RT-PCR assays for MRAS mRNA show that these transcripts are only present in S462 cells. (D) Cluster analysis comparing the relatedness of the R-Ras and classic Ras protein subfamilies. (E) Expression of HA-tagged DN R-Ras in stably transfected ST88-14 cells with varying concentrations of doxycycline. Expression is maximally induced with 2 µg/ml doxycycline. (F) Relative expression levels of HA-tagged DN R-Ras in ST88-14 cells at various time points after addition of 2µg/ml doxycycline. Expression is maximally induced 48hr after the addition of doxycycline.