Skip to main content
. 2014 Jun 18;8(8):1532–1547. doi: 10.1016/j.molonc.2014.06.006

Figure 4.

Figure 4

EMT in HH cells mediated by TGFβ/SMAD signaling: A) Role of β‐catenin, TGFβ and SNAIL was evaluated in MDA‐MB‐231 and HH cells by treating the cells with TGFβ. B) Role of TGFβ was evaluated using transfection of dominant negative TGFβ receptor in MDA‐MB‐231 (HH) cells. The control and transfected cells were treated with TGFβ and effects were compared by western blot technique. C) The nuclear localization of SMADs, SNAIL and β‐catenin was evaluated (i) MDA‐MB‐231 and HH cells were fractionated into cytosolic and nuclear fractions. The proteins were analyzed using western blot techqniue. MDA‐MB‐231 and HH cells were immunostained for (ii) SNAIL and (iii) pSMAD4 (T277). The red staining shows SNAIL or pSMAD4 expression and blue represents nuclear stain C) Binding of SMAD4 to SMAD2/3 was compared in MDA‐MB‐231 with HH cells using immunoprecipitation assay. D) Chromatin immunoprecipitation (CHIP) assay was performed to evaluate SMAD4 DNA binding in MDA‐MB‐231 and HH cells.