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

Figure 3.

Figure 3

Tolytoxin specifically decreases the number of TNT connected cells (a) Rhodamine phalloidin (for actin) and WGA (for membrane) staining in untreated control cells and 3 nM methanol or 3 nM tolytoxin treated SW13 cells. Arrows are indicating the TNTs not touching to the substrate (note that a representative image was selected in a non-TNT containing area since such areas were more pronounced with tolytoxin treatment). Scale bar: 10 μm. (b) Graph showing the percentage of TNT connected cells in Control, 3 nM Me-control or 3 nM tolytoxin treated cells. Data were collected from three independent experiments, 60–80 cells were analysed per condition (p = 0.937 for Control versus 3 nM Me-control, p = 0.0038 for Control versus 3 nM tolytoxin, p = 0.0013 for 3 nM Me-control versus 3 nM tolytoxin. Mean percentage of TNT connected cells in Control: 48.37% ± 4.17, Me-control: 50.33% ± 3.60, 3 nM tolytoxin: 29.23% ± 4.37). (c) Measurement of SW13 cell perimeter in Control, Me-Control and 3 nM tolytoxin treated cells from three independent experiments (40 cells were analysed per condition). Mean perimeter in Control: 1324 ± 42, Me-control: 1318 ± 39, 15 nM tolytoxin 1360 ± 49 (p = 0.995 for control versus 3 nM Me-control, p = 0.827 for control versus 3 nM tolytoxin p = 0.769 for 3 nM Me-Control control versus 3 nM tolytoxin). (d) Actin and microtubule cytoskeleton labeling with rhodamin phalloidin and beta-tubulin respectively in parallel to nuclear DAPI labeling in Control, 15 nM methanol and 15 nM tolytoxin treated SH-SY5Y cells. Arrows indicate TNTs. Scale bar: 10 μm. (e) Graph showing the percentage of TNT connected cells in Control, 15 nM Me-control or 15 nM tolytoxin treated cells from three independent experiments, 60–80 cells were analysed per condition (p = 0.6702 for Control versus 15 nM Me-control, p = 0.0073 for Control versus 15 nM tolytoxin, p = 0.0009 for 15 nM Me-Control versus 15 nM tolytoxin). Mean percentage of TNT connected cells in Control: 43.16% ± 42.15, Me-control: 46.37% ± 2.82, 15 nM tolytoxin: 32.53% ± 2.49. (f) Measurement of cell perimeter in Control, Me-Control and 15 nM tolytoxin treated cells from three independent experiments (40 cells were analysed per condition). Mean perimeter in Control: 899.1 ± 30.42, Me-control: 869.5 ± 45.31, 15 nM tolytoxin: 841.2 ± 62.88; (p = 0.873 for control versus 15 nM Me-control, p = 0.676 for control versus 15 nM tolytoxin, p = 0.906 for 15 nM Me-control versus 15 nM tolytoxin).