Fig. 6

Analysis of EVs after RAB7 overexpression in A2780 CIS cells. A A2780 CIS cells were transfected with empty vector (Mock) and with HA-RAB7, and EVs were purified by immunoisolation. Expression levels of CD9, CD81, ATP5A, and SDHB were evaluated by Western blot in transfected cells, and in EVs purified from these cells. B, C Protein relative abundance was determined through densitometric analysis normalizing against HSP90 protein. D EVs from A2780 CIS were incubated with proteinase K where indicated in the presence or absence of Triton-X100 and then were subjected to immunoblot analysis using antibodies against Alix, TSG101 (as controls), and RAB7. E A2780 CIS cells transfected with empty vector (Mock) and with HA-RAB7 plasmid were treated with CDDP. Subsequently, an MTT assay was performed and cell viability of HA-RAB7-transfected A2780 CIS cells was determined using the matched mock for each concentration as control. F EC50 was determined in A2780 CIS cells treated as indicated after 48 h of incubation with CDDP using the MTT assay and calculated with GraphPad Prism. Data represent the mean ± SEM of at least three independent experiments. * p < 0.05; ** p < 0.01; ***p < 0.001