Journal: eLife

Article Title: Endosomal-lysosomal organellar assembly (ELYSA) structures coordinate lysosomal degradation systems through mammalian oocyte-to-embryo transition

doi: 10.7554/elife.99358

Figure Lengend Snippet: Figure 6. Assembly of endosomal-lysosomal organellar assembly (ELYSA) during migration and limited interaction with the acidification machinery. (A) Snapshots of a time-lapse observation for EGFP fluorescence of germinal vesicle (GV) oocytes injected with Lamp1-EGFP and V1A-mCherry mRNA are indicated. Maximum intensity projection of confocal images with a height of 40 μm (bottom half of the oocyte) are shown. Magnified perinuclear regions (right) are indicated by yellow boxes. Arrowheads indicate assemblies that adhere to each other in the subsequent frame. (B) Maximum intensity projection of deconvolved confocal images at an axial scan range of 80 µm is shown as Z-projection images. Magnified regions are indicated by yellow boxes (a, b) and the LAMP1-EGFP organelle regions are indicated by yellow dotted line.

Article Snippet: The amplicon was then subcloned into the entry vector pDONR221 and transferred into the destination vectors pDEST- CMV- C- EGFP (Agrotis et al., 2019) (a gift from Robin Ketteler [Addgene 0 200 400 600 800 1000 1200 0 1000 2000 3000 4000 5000 6000 7000 8000 9000 10000 11000 0 3 5 8 10 13 15 18 20 23 25 28 30 33 35 38 40 43 45 48 LysoSensor Magic Red 0 200 400 600 800 1000 1200 0 1000 2000 3000 4000 5000 6000 7000 8000 9000 0 2 4 6 9 11131517192224262830323537394143 LysoSensor Magic Red A GV (nuclear) media Isolated punctaAssembly(perinuclear) Assembly In te ns i es Distance from origin (μm) GV MII Isolated puncta media media Assembly (cor cal) Assembly (cor cal) In te ns i es MIIGV BF Ly so Se ns or M ag ic Re d B M er ge BF 20 μm B Satouh et al. eLife 2024;13:RP99358.

Techniques: Migration, Fluorescence, Injection