Fig. 5

Sunitinib-induced EC senescence resulted in vessel leakage and TC extravasation. a The HUVECs treated with vehicle or sunitinib were immunofluorescent stained for VEC (green). b VEC expression was quantified using 10 randomly chosen fields from two experiments using ImagePro Plus (the mean level in the vehicle group was set to 1.0; n = 10). c HUVECs treated with vehicle or sunitinib were subjected to WB assay (means ± SD; n = 3). d The Miles assay was performed as described in the “Materials and methods” section. The Evans Blue (EB) leakage (OD 630 nm) was quantified (n = 10 mice). e Bronchus bronchoalveolar lavage assay to examine the permeability of the lungs from mice treated with vehicle or sunitinib. f, g Balb/c female mice were treated with sunitinib (100 mg/kg/day) for 2 weeks and inoculated with 1 × 10⁶ EGFP-transfected 4T1 cells. The number of green fluorescence-positive CTCs in the blood of tumor-bearing mice was counted using flow cytometry, and data were presented as scatter plots with mean ± SEM (n = 8). Significance was assessed by the Mann-Whitney test