Fig. 5
Identification of ETA as the novel regulator of sEV biogenesis. a Measurements of the number of sEV secreted (left) and the amounts of sEV proteins (right) from WT, ETA-overexpressed (ETA-OE), and ETA knockdown (K/D) MDA-MB231 cells. n = 3. b Immunoblot of various proteins in ETA-OE and ETA-K/D MDA-MB231 cells. n = 3. c Measurement of the number of sEV secreted and the amount of sEV proteins from ETA agonist- or antagonist-treated MDA-MB231 cells. n = 3. d Measurement of the number of sEV and the amounts of sEV proteins from MDA-MB231 cells treated with 10 nM ET1 or 10 nM ET2 in the presence of 100 μM SFX. n = 3. e Measurement of soluble proteins secreted from MDA-MB231 cells in the presence of PD156707 or zibotentan for 24 h. The unit of IL8, IL2, CXCL13, CCL2, TGFB1, and IL6 is pg ml−1. The unit of CX3CL1 is ng ml−1. n = 3. f Tumor mass volume of in BALB/c nude female mice inoculated with MDA-MB231 or MDA-MB231 ETA K/D. Bar, 1 cm. n = 8 per group. g Left top, immunoblot of human CD63 in circulating cancer-cell-derived sEV from MDA-MB231 or MDA-MB231 ETA K/D-bearing mice (n = 8 per group) where similar amounts of protein/lane were verified by Ponceau S staining (Bottom). Right, densitometric analysis of the relative protein intensity of CD63. Experiments were performed with 95% confluent cells. Significance was determined using an unpaired two-tailed Student’s t-test. ***p < 0.001, **p < 0.005 and *p < 0.05. Error bar, SD. Source data (a, c–g) are provided as a Source Data file
