Fig. 7
Modulation of TGF-β pathway activity in response to tepotinib and irradiation alone or in combination. Syngeneic mice were intracranially implanted with GL-261 glioma cells and treated daily with 100 mg/kg tepotinib from day 6 on or solvent, or with a single dose of 10 Gy on day 7, or in combination. RNA and protein were extracted from the right tumor-bearing hemispheres and analyzed for TGF-β pathway activity. A,B. TGF-β1, TGF-β2, TGF-β3, PAI-1 or PDGF-B mRNA expression levels relative to HPRT1 levels were assessed by RT-PCR (*** p < 0.0001, versus control; ### p < 0.0001, versus tepotinib; +++ p < 0.0001, versus radiotherapy). C. pSMAD2 protein levels were assessed by immunoblot. The intensities of protein bands relative to actin were quantified using ImageJ Gel Analysis. D. The cells were untreated or exposed to irradiation in the absence or presence of tepotinib as pre (8 h)- and co-treatment (100 nM), and 24 h later assessed for expression of TGF-β1, TGF-β2, TGF-β3, PAI-1 and PDGF-B mRNA levels. Data are presented as mean ± SD (***, P < 0.001, compared to un-irradiated control)