Figure 5

Preferential exchange of mitochondria from eGFP-E4⁺ECs to MCF-7. A , confocal imaging of co-cultures between MCF7 and eGFP-E4⁺ECs. Prior to co-culture, eGFP-E4⁺ECs were tagged with MitoTracker Deep Red. After 48 hours, co-cultures were stained with WGA AF594 to reveal cell membrane. eGFP-E4⁺ECs mitochondria could be detected in MCF7 (dotted squares). Scale bar: 20 μm. B , Similar experiments were performed with M-orange-MSCs. No exchange was detected at ratio 1/1. Scale bar: 20 μm. C , Tri-culture of MCF7, MSCs (stained with MitoTracker Green) and non-GFP E4⁺ECs (stained with MitoTracker Red) were performed. After 48 hours, cultures were stained with WGA-AF594. E4⁺ECs mitochondria (blue) were detected in MCF7 and MSCs, while MSCs mitochondria (green) remained restricted to MSCs in this set-up. Scale bar: 20 μm. D , Flow cytometry plot of different ratio of co-culture of eGFP-E4⁺ECs/MCF7. Prior to co-culture eGFP-E4⁺ECs were stained with MitoTracker DeepRed. The number of MCF7 up-taking mitochondria from eGFP-E4⁺ECs increased in relation to the number of eGFP-E4⁺ECs. E, graphical representations of the flow cytometry findings.