Fig. 1. Kinase inhibitor screening identifies agents that induce neurite-like process outgrowth in U87 GBM cells.

A Step 1: U87 GBM cells were seeded (2 × 10³ per well) into 96-well plates using the Biomek 4000 automatic workstation. Following incubation for 24 h, cells were treated with 1 μM of the selected KIs (n = 47). Step 2: After 24 h phase-contrast microscopy images were taken at 20x mag using the Olympus IX71 microscope (Micromanager software). Step 3: The length of long thin extensions termed ‘neurite-like process’ were analysed, using the Fiji plugin ‘simple neurite tracer’ (Image J) and the average length neurite-like process per cell determined. B U87 GBM cells were treated for 24 h with 1 μM 47 kinase inhibitors. The average length of neurite-like process was measured and normalised by cell number. The mean ± SD of n = 3 independent experiments is shown. Grey circles vehicle control, black circles not significant (NS) p ≥ 0.05, pink triangles p ≤ 0.05, blue triangles p ≤ 0.01, red triangles p ≤ 0.001 (two-tailed t-test). Kinase inhibitor ‘hits’ from the screening p ≤ 0.001 are indicated with black arrows and include; LY2835219 (CDK4/6), pelitinib (EGFR), CP-673451 (PDGF-Rα/β) and pacritinib (JAK2/FLT3).