Figure 2. Localization of lipids and Chn2 in cysts.

FRET images and quantitative analysis of PI(3,4,5)P₃ (A), PI(3,4)P₂ (B), PI(4,5)P₂ (C) and diacylglycerol (D). The calculated FRET efficiency was colored and shown as in Fig. 1A.The white arrowhead in (D) indicates accumulation of diacylglycerol at the apical membrane. * P<0.05 and ** P<0.01 (E) Schematic view of α1- and β2-Chimaerin encoding 334 and 468 amino acids (AA). SH2 denotes the Src Homology 2 domain. (F) MDCK cysts expressing both dKeima and GFP-β2-Chimaerin were processed and imaged as described in Methods. Localization of GFP-β2-Chimaerin shown in GFP channel (left), dKeima channel (middle) and relative localization of GFP-β2-chimaerin over dKeima (right) shown in pseudocolor mode (at the bottom). White arrowheads indicate apical enrichment of GFP-β2-chimaerin. Scale Bar = 5 µm. (G) Results of quantitative PCR from cysts in early (2 days) and late (10 days) stages. Shown are the averages from three independent experiments with SD.