Inhibitory effect of FGFR signaling on the IFNγ signaling pathway in human cancer cells. A–C, Western blot analysis in MFE280 human endometrial cancer cells using the indicated antibodies; GAPDH was included as a loading control. A, MFE280 cells were starved in medium containing 0.1% FBS for 18 hours, and then treated first with lenvatinib at 1 or 3 μmol/L or with E7090 at 0.3 μmol/L for 1 hour and finally with bFGF (10 ng/mL) for 5 minutes. B, MFE280 cells were pretreated with lenvatinib at 1 or 3 μmol/L for 1 hour and then treated with bFGF (10 ng/mL) for 23 hours. Finally, cells were stimulated with IFNγ (5 ng/mL) for 24 hours. C, MFE280 cells were treated with lenvatinib at 1 or 3 μmol/L for 24 hours and then stimulated with IFNγ (5 ng/mL) for 24 hours. D and E, CXCL10 levels in culture supernatants were analyzed by ELISA. Data are presented as means + SEM of three independent experiments. D, Human renal cell carcinoma 786-O cells were treated first with lenvatinib at 1 or 3 μmol/L or with E7090 at 0.3 μmol/L for 1 hour and then with bFGF (10 ng/mL) for 23 hours. Finally, cells were stimulated with IFNγ (5 ng/mL) for 24 hours. *, P < 0.05; **, P < 0.01, unpaired t test versus bFGF+IFNγ-treated group (blue bar); ###, P < 0.001, Dunnett multiple-comparison test versus bFGF+IFNγ-treated group (blue bar). E, Human hepatocellular carcinoma HuH7 cells were treated first with lenvatinib at 1 or 3 μmol/L or with E7090 at 0.3 μmol/L for 24 hours and then stimulated with IFNγ (5 ng/mL) for 24 hours. ***, P < 0.001, unpaired t test versus IFNγ-treated group (red bar). ##, P < 0.01; ###, P < 0.001, Dunnett multiple comparison test versus IFNγ-treated group (red bar). LEN, lenvatinib; N.D., not determined (i.e., below the lower limit of detection).