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. 2007 Jun 22;93(8):2900–2910. doi: 10.1529/biophysj.106.102582

FIGURE 5.

FIGURE 5

Comparison of 1D and 2D CWT analyses of plasma membrane microdomains of cAMP production stimulated by forskolin (10 μM) in COS1 cells expressing a plasma membrane-targeted FRET-based cAMP reporter. (A) Corrected FRET images of the before forskolin application (left panel) and 2 min (middle panel) and 3 min (right panel) after forskolin treatment. Arrows point to magnified regions containing the plasma membrane microdomains of cAMP production (white boxes). The color bar represents corrected FRET values. (B) 2D wavelet coefficients plot of the plasma membrane region under control conditions (left panel) and normalized subtraction of squared 2D wavelet coefficient matrices (see Materials and Methods) after 2 min (middle panel) and 3 min (right panel) of forskolin application from the control matrix. The color bar represents the difference between squared coefficient matrices normalized to the maximal value. Potential microdomains of cAMP production are visualized by large values. Plasma membrane regions showing statistically significant changes are indicated by circles. (C) Normalized subtraction of squared 1D wavelet coefficient matrices (see Materials and Methods) after 2 min (left panel) and 3 min (right panel) of forskolin application from the control matrix. Plasma membrane microdomains identified by 1D CWT coincide with microdomains identified by 2D analysis (circled region on the left panel C coincides with the region in the bottom of the images presented on panel B; and the circled region on the right panel C coincides with the region in the top of the images presented in panel B). Scale bar is 2 μm.