Supplemental Data

 

Supplemental Data for:
Reese et al. Molecular Cell 6, pp. 4445-455
 
 

Download

Legend

QuickTime video (4.0 MB)

Video 1:  In vivo visualization of PIE-1:GFP dynamics using time lapse fluorescence microscopy. In this time lapse movie of PIE-1:GFP, both fluorescent (top) and Nomarski images (bottom) were collected for each time point and displayed together. In oocytes and newly fertilized embryos, PIE-1:GFP was present uniformly throughout the cytoplasm. In the late one-cell stage, after the pronuclei have formed, PIE-1:GFP levels began to decrease in the anterior cytoplasm and increase in the posterior cytoplasm.  By pronuclear meeting, PIE-1:GFP was found predominantly in the posterior region of the embryo (to the right in this movie). During mitosis, PIE-1:GFP also accumulated on both centrosomes with higher levels on the posterior centrosome. As a result of this asymmetric enrichment, most of the PIE-1:GFP was inherited by the posterior blastomere P1 during the first cleavage.  A similar pattern was observed at each of the subsequent divisions of the germline blastomeres except that PIE-1:GFP became increasingly more nuclear during each interphase.  After cytokinesis, most PIE-1:GFP was found in the germline daughter with only low levels left in the somatic daughter (e.g., EMS in the 4-cell stage).  PIE-1:GFP fluorescence diminished progressively in that cell and was not detected in its progeny.  These observations suggest that PIE-1 segregation to the germ lineage involves mechanisms that act both before cell division (to enrich PIE-1 on the germline side of the cell) and after cell division (to eliminate any PIE-1 remaining in the somatic daughter).

• Summary of this Article
• Full Text of this Article