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. 2014 Nov 25;33(24):2997–3011. doi: 10.15252/embj.201490230

Figure 1. βPix binds Yap/Taz and regulates Yap/Taz localization and transcriptional activity.

Figure 1

  1. Detection of TAZ binding partners using LUMIER assay. The interaction of luciferase-tagged TAZ with individual Flag-tagged proteins was assessed by luciferase assay. Each Flag-tagged protein tested in the screen is represented as a vertical bar, and the median luminescence intensity ratio (mLIR), which reflects the intensity of the interaction with TAZ, is represented by the yellow tone. Several interacting partners are marked.
  2. βPIX interacts with YAP and TAZ. Lysates from HEK293T cells co-transfected with Flag- or HA-tagged βPIX, YAP or TAZ as indicated were subjected to anti-Flag immunoprecipitation (α-Flag IP) and the presence of an associated protein detected by anti-HA immunoblotting. Equivalent protein expression levels were confirmed (Totals).
  3. βPix regulates Yap/Taz localization in polarized epithelial cells. EpH4 cells, transfected with control siRNA or siRNA targeting βPix, were plated at low or high cell densities. After 48 h, cells were fixed and Yap/Taz localization was visualized by confocal immunofluorescence microscopy. Scale bar, 25 μm.
  4. βPix regulates Yap/Taz transcriptional activity. EpH4 cells were transfected with siControl (siCtl) or siβPix, and RNA was extracted at 48 h. A representative experiment showing the relative expression of the Yap/Taz target genes Ctgf and Ankrd1, and the βPix knockdown efficiency as determined by qPCR is plotted as the mean ± the range.

Source data are available online for this figure.