Fig. 7.

The relationship between Ama and Sprouty is conserved in human glioblastoma cell lines. (A) Immunoblot showing LSAMP, SPRY2 and GAPDH levels in U251 cells stably expressing non-targeting shRNA (NT) or LSAMP shRNA (LSAMPsh). Knockdown of LSAMP increases SPRY2, with relative densitometry levels for each target labeled below. (B) Immunoblot showing LSAMP, SPRY2 and vinculin levels in T98G cells expressing a doxycycline-inducible shRNA. Cells were treated for 3 days with the corresponding amounts of doxycycline (ng/ml in 0.15% DMSO) before harvesting. Dose-dependent knockdown of LSAMP gradually increases SPRY2. Relative densitometry levels for each target are labeled below. Results in A and B are representative of three independent experiments. (C) Heatmap displaying mRNA expression z-score (RNA Seq V2 RSEM) of LSAMP and SPRY2 in each row in GBM human patients (TCGA PanCancer Atlas) showing a significant inverse correlation between those genes. Each column represents a tumor tissue sample. The q-value was generated from a two-sample t-test. (D) Kaplan–Meier plot revealing that GBM patients (TCGA PanCancer Atlas) with EGFR mutations have a significant decrease in survivability with high levels of LSAMP mRNA. The P-value was generated from a logrank test. (E) Kaplan–Meier plot of GBM patients with EGFR amplifications reveals that patients with high LSAMP mRNA have a significant decrease in survivability. The P-value was generated using a logrank test.