Fig. 2. Accumulation of membrane vesicles in cells expressing Gas3/PMP22. (A) Time-lapse images of SCs overexpressing Gas3/PMP22-GFP. Frames at selected hours after microinjection, as indicated, showing Schwann cells overexpressing Gas3/PMP22-GFP. Arrowhead indicates cluster of vesicles in the perinuclear region and asterisks indicate vacuoles. Bar, 18 µm. (B) Time-lapse images of NIH-3T3 cells overexpressing Gas3/PMP22-GFP. Frames at selected hours after microinjection, as indicated, of a representative cell injected with pEGFP-N1-Gas3/PMP22 (10 ng/µl) are shown. Arrowhead indicates cluster of vesicles in the perinuclear region; arrows indicate vacuoles; asterisks indicate a vacuole close to the area retracted from the adhesion substrate. Bar, 14 µm. (C) Electron micrograph of a SC from sciatic nerve of homozygous PMP22_tg rat, 100,000x. Intracellular myelin figures not associated with an axon are present (arrow). Moreover, empty vacuoles made up by redundant layers of SC membrane (asterisk) and plasma membrane invagination (arrowhead) can be observed. (D) Electron micrograph of a homozygous PMP22_tg SC in culture (5 days), 60,000x. Concentric sheaths of membrane are present in the cytoplasm (arrows). Vacuoles can be observed either in the centre of the layer, or related to the SC membrane (asterisk).

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