Western Blot Analysis

Ba/F3 cells and patient derived cell lines were treated for 6 hours with lorlatinib, LA7 or LA9 unless otherwise noted. Total protein lysates were analyzed by western blotting with the following antibodies and dilutions. (all from Cell Signaling Technology): phospho-ALK Y1604 (3341, 1:1000), phospho-ALK Y1282/83 (9687, 1:1000), ALK (3633, 1:1000), phospho-AKT S473 (4060, 1:1000), AKT (4691, 1:1000), phospho-ERK1/2 T202/Y204 (9101, 1:1000), ERK1/2 (9102, 1:1000), phospho-S6 S240/244 (5364, 1:10,000), S6 (2217, 1:10,000), and β-Actin (4970, 1:1000).