Fig. 4
From: Highly-purified rapidly expanding clones, RECs, are superior for functional-mitochondrial transfer
MSC/REC connect with ρ0 cells through F-actin containing microtubules and gap junction. Representative immunofluorescent images of (A) ρ0 A549 co-culture with MSC, (B) ρ0 A549 co-culture with REC, (C) ρ0 HeLa co-culture with MSC, and (D) ρ0 HeLa co-culture with REC. MSC/REC (red) were labeled with Mito Tracker Deep Red, ρ0 cells were labeled with DAPI (blue), and all cells were counterstained with phalloidin (F-actin, green). White arrows depict microtubules connecting MSC/REC and ρ0 cells. E Cytochalasin D was an inhibitor of actin formation, added during ρ0 A549 co-culture with MSC (I) and REC (II), respectively. Cytochalasin D was added during ρ0 HeLa co-culture with MSC (III) and REC (IV), respectively. MSC/REC (green) were labeled with Mito Tracker Green, ρ0 cells were labeled with DAPI (blue), and all cells were counterstained with phalloidin (F-actin, red). F Mitochondria in TNTs were observed by transmission electron microscopy. G Mitochondria in microvesicles were observed by transmission electron microscopy. Representative immunofluorescent images of (H) ρ0 A549 co-culture with MSC (I) and REC (II), respectively. (III–IV) ρ0 HeLa co-culture with MSC (III) and REC (IV), respectively. MSC/REC (red) were labeled with Mito Tracker Deep Red and all cells were counterstained with connexin 43 (Gap junction, green). The white arrow shows MSC/REC generating gap junction to transfer mitochondria. I When exposed to Cytochalasin D, the number of TNTs in the ρ0 A549 cell line and ρ0 HeLa cell line was calculated. J Quantitative analysis of the relative fluorescence intensity of Cx43 protein produced by ρ0 A549 cells co-cultured with MSC and REC, respectively. K Quantitative analysis of the relative fluorescence intensity of Cx43 protein produced by ρ0 HeLa cells co-cultured with MSC and REC, respectively. Data were based on three independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001