XB-IMG-144529
Xenbase Image ID: 144529
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Figure S2. Quantification of Xdsh-GFP Membrane Association
To quantify the degree of membrane association of Xdsh-GFP, we
calculated its colocalization with membrane-tethered RFP
(memRFP [S9]). The plot in this figure shows the ratio of pixels in
which GFP and RFP signals are colocalized to the total pixels. A
threshold is applied to remove low-intensity signal that is likely
to be nonspecific. This ratio, the “colocalization coefficient,” was
attained with Zeiss LSM5 software.
First, as a control, we measured colocalization of Xdsh-GFP and
memRFP in control animal caps (column 1). Expression of unlabeled
Dvl-KM-caax resulted in a dramatic increase in Xdsh-GFP colocalization
with memRFP (column 2), demonstrating the efficacy of this
quantification method (see also Figures 1D and 1F in the main text).
We next quantified colocalization in DMZ cells. As compared to
normal animal caps cells, DMZ cells display a much higher fraction
of Xdsh-GFP colocalized with memRFP (column 3). However, in
DMZ cells expressing unlabeled Xdsh-MA, we observe as much as
a 10-fold reduction in the colocalization of Xdsh-GFP and memRFP
(column 4). These data indicate that Xdsh-MA effectively removes
wild-type Dvl from the cell membrane in cells undergoing convergent
extension. Image published in: Park TJ et al. (2005) Copyright © 2005. Image reproduced with permission of the Publisher, Elsevier B. V. Larger Image Printer Friendly View |