Xenbase Image ID: 138556

Figure 2. Notum Causes Wnt Deacylation and Inactivation (A) The Wnt3a protein modified by Notum became hydrophilic in the Triton X-114 detergent-aqueous phase separation assay. Wnt3a from mock or Notum(S239A)-expressing cells was hydrophobic and partitioned in the detergent (De) phase, but Wnt3a from Notum- or Notum-TM-expressing cells partitioned in the aqueous (Aq) phase; total input, T. (B) Wnt3a CM from Notum- or Notum-TM-expressing cells was inactive and induced minimal phosphorylation of LRP6 and Dvl2 in mouse embryonic fibroblast cells, whereas Wnt3a CM from mock, Notum(S239A)-, or Notum(S239A)-TM-expressing cells induced phosphorylation of LRP6 and Dvl2. Note that Wnt3a from Notum- or Notum-TM-expressing cells exhibited slightly faster migration. WCL, whole cell lysates. (C) Wnt3a CM from Notum or Notum-TM-expressing cells induced minimal TOPFLASH in HEK293T cells, whereas Wnt3a CM from mock, Notum(S239A)-, or Notum(S239A)-TM-expressing cells induced TOPFLASH. Error bars represent SD of triplicated experiments. (D) Wnt3a secreted from Notum-expressing cells exhibited minimal binding to mFz8CRD-IgG, whereas Wnt3a secreted from mock or Notum(S239A)-expressing cells exhibited binding. (E) Notum, but not Notum(S239A), reduced Wnt3a acylation when they were co-expressed in HEK293T cells. See also Figure S2. Image published in: Zhang X et al. (2015) Copyright © 2015. Image reproduced with permission of the Publisher, Elsevier B. V. Larger Image Printer Friendly View

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