Figure 1: TGFβ induces nuclear accumulation of TβRI intracellular domain (TβRI-ICD).
From: TRAF6 ubiquitinates TGFβ type I receptor to promote its cleavage and nuclear translocation in cancer

(a, b) Knockdown of TβRI by siRNA was performed to demonstrate the specificity of the V22 and H100 antibodies, which recognize the intracellular and extracellular domain of TβRI, respectively, in immunofluorescence (a; DAPI was used to visualize cell nuclei, scale bar 20 μm). (b) Cell lysates of PC-3U cells treated or not with TGFβ, were fractionated into cytoplasmic and nuclear proteins and subjected to SDS–gel electrophoresis, followed by immunoblotting using V22 and H100 antibodies. β-tubulin and lamin A served as controls for the cytoplasmic and nuclear fractions, respectively. Molecular weight markers are indicated. (c) Representative confocal microscopy pictures of C-terminally pEGFP-N3-tagged wt TβRI (GFP- TβRI) and pEGFP-N3-vector (GFP) expressed in PC-3U cells treated with TGFβ for 6 h are shown (left panel) and quantified in right panel (mean±s.d., n=3–5 independent experiments where N=350 cells where counted in each group, *P<0.0002, ANOVA). Cell nuclei were stained with DAPI. Scale bar 20 μm.