Figure 7: TβRI nuclear localization in human breast and lung carcinoma cells is associated with tumour invasion.
From: TRAF6 ubiquitinates TGFβ type I receptor to promote its cleavage and nuclear translocation in cancer
(a) Human breast carcinoma (MDA-MB-231) cells were starved and treated with TGFβ in the presence or absence of the PKCζ pseudosubstrate (P.S.) or TAPI-2, as indicated. Endogenous C-terminal TβRI was visualized by immunofluorescence using the V22 antibody (TRITC). Quantification of the number of cells showing endogenous TβRI in the nucleus is shown on the right side of the panel (mean±s.d., n=3 independent experiment, where N=200–300 cells where counted in each group, *P<0.001, ANOVA). Scale bar 20 μm. (b) Invasion assay for MDA-MB-231 treated with TGFβ in the absence or presence of TAPI-2 and PKCζ P.S. Cells were visualized by staining with crystal violet cell stain solution. Right panel presents mean values for optical density (OD) of invasive cells. Error bars represents s.d. (n=3 independent experiments, *P<0.05; Students t-test). (c) Human lung carcinoma (A549) cell were starved and treated with TGFβ in the presence or absence of the PKCζ P.S or TAPI-2, as indicated. Endogenous C-terminal TβRI was visualized by immunofluorescence using the V22 antibody (TRITC). Quantification of the number of cells showing endogenous TβRI in the nucleus is shown on the right side of the panel (mean±s.d., n=3 independent experiment, where N=200-300 cells where counted in each group, *P<0.0003, ANOVA). Scale bar 20 μm. (d) Invasion assay for A549 cells treated with TGFβ in the absence or presence of TAPI-2 and PKCζ P.S. Cells were visualized by staining with crystal violet cell stain solution. Right panel presents mean values for OD of invasive cells. Error bars represents mean±s.d. (n=3 independent experiments; *P<0.05, **P<0.002, ANOVA).
