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Supplementary Figure 2
(A) Co-immunoprecipitation of overexpressed proteins shows a weak interaction of Pgam5 with Arrb2, which is
slightly increased by co-expression of Dvl2. (B) Western blot corresponding to Fig. 1D. The binding site of Pgam5
to Dvl2 was mapped to the region interspacing the PDZ and DEP domain of Dvl2 by co-immunoprecipitation of
Pgam5-HA with a series of truncated Dvl2-Myc constructs (1-8) as illustrated.
(C) Overexpression of PGAM5 partially reverted WNT3A niduced electrophoretic mobility shifts of endogenous
DVL2 in HEK 293T cells, indicating that PGAM5 dephosphorylates DVL2. (D) HA-DVL2 was immunoprecipitated,
washed with high stringency to remove bound proteins and incubated with recombinant GST, GST-PGAM5δ
(lacking the N-terminal mitochondrial targeting sequence) or Calf Intestinal Alkaline Phosphatase (CIP) for 30
min at 37°C. The sample incubated with GST showed two bands. After incubation with GST-PGAM5δ or CIP
only one band with higher electrophoretic mobility remained, which confirmed that DVL2 is directly
dephosphorylated by PGAM5.
Image published in: Rauschenberger V et al. (2017)
Copyright © 2017. Image reproduced with permission of the publisher and the copyright holder. This is an Open Access article distributed under the terms of the Creative Commons Attribution License.
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