Figure 2. Anti-EMT function of SOCS1: EMT markers were down-regulated in cells transduced with SOCS1, while promoted upon SOCS1 knock-down.

HA & HA-SOCS1, and sh & sh-SOCS1 HCT116 p53+/+ cells were established by transfection of respective vectors by electroporation as described in Materials and Methods. Cells maintained in the selection media were treated with H₂O₂ (300 μM for 2 h) and changes in EMT markers were analyzed by immunoblotting. A representative blot is shown for each cell system A. Densitometric analysis of immunoblots normalized to beta-actin showing relative expression of EMT regulators. Data represent mean+SE obtained from 3 independent experiments B. HCT116 p53 +/+ HA & HA-SOCS1 cells were treated with H₂O₂ as above and then changes in EMT markers and SOCS1 location were examined by immunofluorescence analysis C. Vimentin and E-cadherin colocalize at the cell membrane in control HA cells shown in the merged figure (white arrows in upper left panel). SOCS1 colocalizes with Twist in the perinuclear region (Lower left panel). In SOCS1-transduced cells E-cadherin expression levels increased over vimentin (Upper right panel), and SOCS1 expression was seen both in the cytosol and nucleus with reduced Twist expression (Lower right panel).