Figure EV5. Apelin inhibition protects from anti‐angiogenic therapy‐induced metastases.

• A
  Number of metastatic lung foci in untreated (control) and sunitinib‐treated (60 mg/kg, three times a week from tumor initiation) NeuT;Apln ^+/+ and NeuT;Apln ^−/− mice with large size‐matched tumors. Data of individual lung sections and means (black bars) are shown. Identified outliers by ROUT method (Q = 1%) were obviated. Right panels show representative H&E images, where black arrows and insets indicate metastatic foci. Scale bars = 1,000 μm (large panels) and 50 μm (insets). ***P < 0.001; one‐way ANOVA; NeuT;Apln ^+/+ control (n = 9), NeuT;Apln ^−/− control (n = 8), NeuT;Apln ^+/+ sunitinib (n = 6), NeuT;Apln ^−/− sunitinib (n = 7); and three sections per lung were analyzed.
• B, C
  Number of metastatic lung foci per lung section from 1 to 1.5 cm³ size‐matched E0771 control (shRenilla) and Apelin‐depleted (shApln) E0771 tumors orthotopically injected into C57BL/6J Apln ^+/+ (B) or orthotopically injected into C57BL/6J Apln ^+/+ or Apln ^−/− mice (C). Mice were either left untreated (control) or treated with sunitinib (60 mg/kg, starting at day 5 after tumor injection, five times a week). Data of individual lung sections and means (black bars) are shown. Right panels show representative H&E images, where black arrows and insets indicate metastatic foci. Scale bars = 1,000 μm (large panels) and 50 μm (insets). E0771 shRenilla control (n = 7), E0771 shApln control (n = 4), E0771 shRenilla sunitinib (n = 5), E0771 shApln sunitinib (n = 3) (B) or Apln ^+/+ shRenilla control (n = 3), Apln ^−/− shApln control (n = 3), Apln ^+/+ shRenilla sunitinib (n = 2), Apln ^−/− shApln sunitinib (n = 3) (C) and three sections per lung were analyzed. **P < 0.01; ***P < 0.001; one‐way ANOVA.