Figure 7
From: The Protease Activated Receptor2 Promotes Rab5a Mediated Generation of Pro-metastatic Microvesicles
MVs from breast cancer patients display higher metastatic potential. Both normal as well as human breast cancer tissue samples were collected and subjected to analysis of Rab5a expression at transcription level (A) and protein level (B), (C) and (E). PAR2 protein expression was also estimated in those tissues (B) and (D). MVs were isolated from the blood of both breast cancer as well as normal individuals and quantified by Bradford assay and western blotting with TF marker (F) and (G). DLS study was performed with these MVs to determine their size (H). Migration (I) and (J) as well as invasive (K) and (L) potential of these blood-borne MVs was analyzed by wound healing assay and transwell invasion assay, respectively. MVs, isolated from the blood samples were fused with MCF-7 cells and p38 MAPK activation was assessed by western blotting (M). MCF-7 cells were treated with SB203580 for an hour followed by the addition of blood-derived MVs. Migration and invasion was checked by wound healing assay and transwell invasion assay, respectively to determine the metastatic potential of MVs (N–Q). The experiments were repeated at least for three times. Data presented over here are as Mean +/− S.E. of the Mean and differences are statistically significant at p < 0.05 using student’s t-test and ‘ns’ represents non-significant differences.
