Supplementary Data from Structural Alterations of MET Trigger Response to MET Kinase Inhibition in Lung Adenocarcinoma Patients

Supplementary Methods Supplementary Figure S1. Characterization of KIF5B- and STARD3NL-MET rearrangements. A) IGV screenshots of next-generation sequencing reads from breakpoint regions of KIF5B- and STARD3NL-MET positive patient tissue. Due to homology in both regions the breakpoint region is marked with a red bar. The spanning read indicates the genomic sequence of the rearrangement. B) Dideoxy sequencing of exonic breakpoint regions of patient cDNA extracted from FFPE cuts for KIF5B- and STARD3NL-MET. C) PET/CT scans of KIF5B-MET patient. Numbers below indicate reduction of SUVmax under crizotinib therapy. D) Immunoblots of NIH-3T3 cells transduced with KIF5B-MET (upper panel) and STARD3NL-MET (lower panel) treated with 1 µM crizotinib or tepotinib over time. HSP90 serves as loading control. A representative blot from n=3 independent experiments is shown. E) Dose-response curves (72 h) of Ba/F3 cells stably transduced with KIF5B-MET or STARD3NL-MET supplemented with IL-3 (10% conditioned Wehi-3b media) and treated with crizotinib, tepotinib and erlotinib (as control compound). (n=3) Supplementary Figure S1. Characterization of KIF5B- and STARD3NL-MET rearrangements. A) IGV screenshots of next-generation sequencing reads from breakpoint regions of KIF5B- and STARD3NL-MET positive patient tissue. Due to homology in both regions the breakpoint region is marked with a red bar. The spanning read indicates the genomic sequence of the rearrangement. B) Dideoxy sequencing of exonic breakpoint regions of patient cDNA extracted from FFPE cuts for KIF5B- and STARD3NL-MET. C) PET/CT scans of KIF5B-MET patient. Numbers below indicate reduction of SUVmax under crizotinib therapy. D) Immunoblots of NIH-3T3 cells transduced with KIF5B-MET (upper panel) and STARD3NL-MET (lower panel) treated with 1 µM crizotinib or tepotinib over time. HSP90 serves as loading control. A representative blot from n=3 independent experiments is shown. E) Dose-response curves (72 h) of Ba/F3 cells stably transduced with KIF5B-MET or STARD3NL-MET supplemented with IL-3 (10% conditioned Wehi-3b media) and treated with crizotinib, tepotinib and erlotinib (as control compound). (n=3)