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Fig. 4 | Journal of Nanobiotechnology

Fig. 4

From: TSPAN4-positive migrasome derived from retinal pigmented epithelium cells contributes to the development of proliferative vitreoretinopathy

Fig. 4

The functional properties of TSPAN4 RPE. RPE were transfected with lentiviral constructs (empty vector as control (Lv-ctrl), or vector overexpressing TSPAN4 (Lv-TSPAN4)). Supernatant of the cultured cells were collected. EVs in the supernatants were isolated by ultracentrifuge. A Nanoparticle Tracking Analysis (NTA) of the EVs from Lv-ctrl and Lv-TSPAN4 groups. Vesicles were counted at each 10 diameters. B transmission electron microscope (TEM) of collected EVs. Scale bar, 50 μm. C Recipient RPE cells were transfected with Lv-mCherry. mCherry+ RPE were incubated with collected EVs isolated from TSPAN4-GFP transfected RPE cells and were visualized by confocal microscope. Sectional image confirmed the existence of TSPAN4+ migrasomes inside the recipient cell. White line demarcated the mCherry+ cell. D, E RPE were transfected with Lv-ctrl or Lv-TSPAN4. The EVs from these two groups were isolated and added to the culture medium of native RPE. The recipient RPE was trypsinized and subjected to Transwell or CCK8 assay. The migration ability was evaluated by calculating the average number of migrated cells in 5 random vision fields (D). Proliferation capability was tested with CCK-8 and measured with OD450 (E)

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