Figure 6
From: The Protease Activated Receptor2 Promotes Rab5a Mediated Generation of Pro-metastatic Microvesicles
MVs Generated from MDA-MB-231 Cells upon Rab5a Activation Induces MCF-7 Cell Metastasis in A p38 Dependent Manner. Comparative TF level in MDA-MB-231 vs MCF-7 cells (A). MVs were isolated from MDA-MB-231 cells after Rab5a transfection and trypsin addition and quantified by western blotting (B). MDA-MVs were incubated with MCF-7 cells and MVs fusion was confirmed by analyzing TF in MCF-7 cells (C). MVs fused MCF-7 cell migration was assessed by wound healing assay (D) and (E). Invasive potential of MVs was also verified by transwell invasion assay after fusing the MVs with a non-invasive cell MCF-7 (F) and (G). MVs were generated from MDA-MB-231 cells after trypsin treatment alongside control and were fused with MCF-7 cells. p38 MAPK phosphorylation was analyzed in these MCF-7 cells by western blotting (H). MCF-7 cells were treated previously with p38 MAPK activity inhibitor, SB203580 followed by MDA-derived MVs addition. Migration and invasion of these MCF-7 cells was determined by wound healing assay and transwell invasion assay respectively (I–L). 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.
