Wnt5a Control of Cell Polarity and Directional Movement by Polarized
Redistribution of Adhesion Receptors
Eric S. Witze, Elizabeth S. Litman, Gretchen M. Argast, Randall T. Moon,
Natalie G. Ahn
Supporting Online Material
This supplement contains:
Materials and Methods
Figs. S1 to S6
References and Notes
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Other Supporting Online Material for this manuscript includes the following:
(available at www.sciencemag.org/cgi/content/full/320/5874/365/DC1)
Movies S1 to S13
Movie s1
Live cell imaging of the WM239A cell shown in Fig. 2A, surface-labeled with
anti-MCAM antibody coupled to AlexaFluor-488. Times are in minutes following Wnt5a
treatment.
Movie s2
Live cell imaging of the cell in Movie S1, membrane-labeled with BODIPY TR
C5-ceramide, as shown in Fig. 2A.
Movie s3
Live cell imaging of WM239A cells, surface-labeled with anti-MCAM antibody
coupled to AlexaFluor-488. Cells were treated under conditions identical to those in
Movie S1.
Movie s4
Live cell imaging of cells shown in Movie S3, membrane-labeled with
BODIPY TR C5-ceramide.
Movie s5
Live cell imaging of a WM239A cell transfected with MCAM-GFP, as shown in
Fig. S3A. Times are in minutes following Wnt5a treatment.
Movie s6
Live cell imaging of the cell shown in Movie S5, transfected with mRFP-actin,
as in Fig. S3B.
Movie s7
WM239A cells transfected with MCAM-GFP and Wnt5a–RNAi, treated
with Wnt5a, and imaged live for 100 min. Times are in minutes following Wnt5a addition.
Cells were scored positive when the accumulation of MCAM-GFP into an asymmetric
structure could clearly be observed. When the process of MCAM-GFP accumulation
could not be observed, cells were not scored. In order to minimize file size, movies were
truncated after MCAM-GFP enrichment had ceased within the 100 min monitoring
9
period. Movies S7, S8, and S9 respectively correspond to Wnt5a-treated cells 2, 3, and
7 in Fig. 2C.
Movie s8
WM239A cells transfected with MCAM-GFP and Wnt5a–RNAi, treated
with Wnt5a, and imaged live for 100 min. Times are in minutes following Wnt5a addition.
Cells were scored positive when the accumulation of MCAM-GFP into an asymmetric
structure could clearly be observed. When the process of MCAM-GFP accumulation
could not be observed, cells were not scored. In order to minimize file size, movies were
truncated after MCAM-GFP enrichment had ceased within the 100 min monitoring
9
period. Movies S7, S8, and S9 respectively correspond to Wnt5a-treated cells 2, 3, and
7 in Fig. 2C.
Movie s9
WM239A cells transfected with MCAM-GFP and Wnt5a–RNAi, treated
with Wnt5a, and imaged live for 100 min. Times are in minutes following Wnt5a addition.
Cells were scored positive when the accumulation of MCAM-GFP into an asymmetric
structure could clearly be observed. When the process of MCAM-GFP accumulation
could not be observed, cells were not scored. In order to minimize file size, movies were
truncated after MCAM-GFP enrichment had ceased within the 100 min monitoring
9
period. Movies S7, S8, and S9 respectively correspond to Wnt5a-treated cells 2, 3, and
7 in Fig. 2C.
Movie s10
WM239A cells transfected with MCAM-GFP and Wnt5a-RNAi, treated
with buffer (minus Wnt5a), and imaged live for 100 min. Times are in minutes following
buffer addition. Cells were scored positive when the accumulation of MCAM-GFP into an
asymmetric structure could clearly be observed. When the process of MCAM-GFP
accumulation could not be observed, cells were not scored. Movies S10, S11, and S12
respectively correspond to control cells 3, 2, and 1 in Fig. 2C.
Movie s11
WM239A cells transfected with MCAM-GFP and Wnt5a-RNAi, treated
with buffer (minus Wnt5a), and imaged live for 100 min. Times are in minutes following
buffer addition. Cells were scored positive when the accumulation of MCAM-GFP into an
asymmetric structure could clearly be observed. When the process of MCAM-GFP
accumulation could not be observed, cells were not scored. Movies S10, S11, and S12
respectively correspond to control cells 3, 2, and 1 in Fig. 2C.
Movie s12
WM239A cells transfected with MCAM-GFP and Wnt5a-RNAi, treated
with buffer (minus Wnt5a), and imaged live for 100 min. Times are in minutes following
buffer addition. Cells were scored positive when the accumulation of MCAM-GFP into an
asymmetric structure could clearly be observed. When the process of MCAM-GFP
accumulation could not be observed, cells were not scored. Movies S10, S11, and S12
respectively correspond to control cells 3, 2, and 1 in Fig. 2C.
Movie s13
Live cell imaging of cells transfected with MCAM-Dendra2, shown in Fig. 4E.
To view these movies, download a QuickTime viewer.
