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Figure 2


Fig. 2. Apical PtdIns(3,4,5)P3 is promoted by high-density culture. (A) PtdIns(3,4,5)P3 distribution in two-cell embryos cultured in high- or low-density conditions. PtdIns(3,4,5)P3 was detected with GFP-PHGRP1 or GFP-PHAkt, and fluorescence linescans (right) were aquired across the apical membrane, avoiding the nucleus (blue and red bars). Cytosolic fluorescence was normalised to 1 and apical PtdIns(3,4,5)P3 was considered to be present when fluorescence in the cortex reached a value of ≥1.2 (dotted line on the linescan graphs). Arrowheads indicate the position of the second polar body. (B) Analysis of the data collected using linescan analysis, as shown in A. The proportion of embryos with (blue) or without (red) apical PtdIns(3,4,5)P3 is expressed as a percentage. Culture conditions and the number of embryos examined are indicated on the x-axis. *P<0.0001; #P<0.001.





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