Figure 4: Identification of the TGFβ-induced cleavage site for TACE in the extracellular domain of TβRI.
From: TRAF6 ubiquitinates TGFβ type I receptor to promote its cleavage and nuclear translocation in cancer
(a) Total cell lysates derived from PC-3U cells transiently transfected with C-terminally HA-tagged wt TβRI or the corresponding G120I mutant were stimulated with TGFβ as indicated, was subjected to immunoblotting for HA to detect the TβRI-ICD fragment. The filter was reprobed with β-actin antibodies to show equal loading of proteins in all lanes, and activation of Smad2, by a phospho-specific antiserum (p-Smad2). The p-Smad2 filter was reblotted with total Smad2. (b) Representative confocal microscopy pictures of PC-3U cells ectopically expressing C-terminally HA-tagged wt TβRI or the corresponding G120I mutant were stimulated with TGFβ for 0.5 h and thereafter stained with HA antibody. Staining with DAPI was used to visualize cell nuclei. Scale bar 20 μm.
