Figure 1 from E2F1-Associated Purine Synthesis Pathway Is a Major Component of the MET-DNA Damage Response Network

Responses of a panel of MET-positive MET-driven and non-MET-driven human cancer cell models to selective MET targeting. A, Whole-cell lysates were subjected to Western blotting using specific antibodies against MET, pMET, and pS6 following the treatment either with vehicle or 50 nmol/L tepotinib (EMD1214063, shortly EMD) for 24 hours. Western blots representative of N = 3 independent experiments are shown. B, Whole-cell lysates were subjected to Western blotting using a specific antibody against p4E-BP1 following the treatment either with vehicle or 50 nmol/L tepotinib (EMD1214063, shortly EMD) for 24 hours. Western blots representative of N = 3 independent experiments are shown. C, EBC-1 (top) and HCC827 (bottom) xenograft tumor volume upon vehicle or METi treatment (N = 10). D, pMET Tyr1234/5 expression in EBC-1 (left) and HCC827 (right) tumor xenografts treated by vehicle or METi (sampling was performed 2 hours posttreatment). Representative images (top) and quantification of pMET-positive area (bottom) are provided [EBC-1: N = 10 (control), N = 10 (tepotinib); HCC827: N = 6 (control), N = 4 (tepotinib); unpaired Student t test, *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001]. E, Representative images (left) of nuclear foci of γH2AX in MET-driven GTL-16 and EBC-1 and non-MET-driven HCC827 and H820 cells. Untreated or METi-treated (50 nmol/L) cells were fixed; γH2AX-positive foci were visualized by confocal microscopy and counted (right), N = 3, unpaired Student t test, *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001.