fig3

Figure 3. In vivo effect of GW4869 treatment on the growth of paclitaxel-resistant PC3-R cells’ xenografts. Male athymic nude mice were treated with vehicle or GW4869, and various study parameters were assessed as described in the methods. (A) Average body weight (mean ± SEM) at the start and day 40 of the study for control mice (n = 3) and GW4869 treated mice (n = 4) mice. (B) Average xenograft volume (mean ± SEM) in control and GW4869 treated mice survived at measured time points is presented. (C) In the end, mice were sacrificed, xenografts were excised, and their weight was measured. Average tumor weight in the control group (n = 6 xenografts) and the GW4869 group (n = 8 xenografts) is presented as mean ± SEM. Unpaired t test was used to calculate the statistical significance, *P = 0.017. (D) CD63 expression in xenograft tissues from the vehicle control (VC) group and GW4869 group (n = 6 each) was measured by IHC. For each image, ten random areas were analyzed for IHC scoring as described in the methods. Mean IHC scores are presented as mean ± SEM in the bar diagram. Representative images are shown. (E) EVs isolated from the plasma of control (n = 3) and GW4869 treated mice (n = 3) were analyzed by NTA, and the size distribution (mean ± SEM) of EVs was plotted as a percentage of total EVs. (F) EVs from VC (n = 3) and GW4869 treated (n = 4) mice were used for the analysis of syntenin, CD63, and GOLGA2 by Western blotting. Representative immunoblots are shown. (G) Densitometry analysis of syntenin and CD63 expression was performed and normalized with corresponding band intensity in Ponceau-stained membrane. The relative band intensity is presented as mean ± SEM.