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. 2019 Apr 5;9:5741. doi: 10.1038/s41598-019-42161-6

Figure 4.

Figure 4

Tolytoxin decreases mitochondria transfer in SW13 and SH-SY5Y cells. (a) Mito-GFP transfected SW13 donor cells in Control, 3 nM Me-control and 3 nM tolytoxin co-cultures (left panel), acceptors transfected with H2B-mCherry (middle panel), donor and acceptor cells in different co-culture conditions (right panel). Arrows indicate mitochondria that had been received by acceptor cells marked with asterisk. Scale bar: 10 μm. (b) Upper graph showing the percentage of mitochondria transfer in each condition. Mean percentage of mitochondria transfer in Control: 11.34% ± 1.91, 3 nM Me-control: 12.99% ± 2.17, 3 nM tolytoxin: 4.84% ± 1.31 (p = 0.80 for Control versus 3 nM Me-control, p = 0.043 for Control versus 3 nM tolytoxin, p = 0.0093 for 3 nM Me-control versus 3 nM tolytoxin). Lower graph showing the average number of mitochondria puncta per acceptor cell in each condition. Mean number of mitochondria puncta in Control: 2.8 ± 0.5, 3 nM Me-control: 2.22 ± 0.42, 3 nM tolytoxin: 1.16 ± 0.11 (p = 0.62 for Control versus 3 nM Me-control, p = 0.19 for Control versus 3 nM tolytoxin, p = 0.48 for 3 nM Me-control versus 3 nM tolytoxin). (c) Mito-DsRed expressing SH-SY5Y donors in Control, 15 nM Me-control and 15 nM tolytoxin co-cultures (left panel), H2B-GFP expressing acceptor cells (middle panel), donor and acceptor cells in different co-culture conditions (right panel). Arrows indicate mitochondria that had been received by acceptor cells marked with asterisk. Scale bar represents 15 μm. (d) Upper graph showing the percentage of mitochondria transfer in each condition. Mean percentage of mitochondria transfer in Control: 56.35% ± 11.3, 15 nM Me-control: 40.51% ± 3.77, 15 nM tolytoxin: 10.37% ± 2.99 (p = 0.188 for Control versus 15 nM Me-control, p = 0.0004 for Control versus 15 nM tolytoxin, p = 0.0069 for 15 nM Me-control versus 15 nM tolytoxin). Lower graph showing the average number of mitochondria puncta per acceptor cells in each condition. Mean number of mitochondria puncta in control: 2.75 ± 0.75, 15 nM Me-control: 1.42 ± 0.2 15 nM tolytoxin: 1.5 ± 0.28 (p = 0.08 for Control versus 15 nM Me-control, p = 0.15 for Control versus 15 nM tolytoxin, p = 0.99 for 15 nM Me-control versus 15 nM tolytoxin). Data were collected from three independent experiments where 60–80 cells were analysed per condition for both cell types.