Fig. 3
From: The E3 ubiquitin ligase NEDD4 mediates cell migration signaling of EGFR in lung cancer cells
NEDD4 does not ubiquitinate and downregulate PTEN. a, NEDD4 was co-expressed with flag-PTEN or Myc-ACK1 by transfection in HEK293 cells. Ubiquitinated ACK1 or PTEN was precipitated with bead-bound GST-Uba from the cell lysates followed by immunoblotting with anti-Myc or anti-flag antibodies. b, Lung cancer A549 cells were infected with lenti-viral vector pLKO.1 or the vector-loaded shNEDD4. NEDD4 in the cell lysates was detected by immunoblotting with anti-NEDD4 (second top panel). The effect of knockdown NEDD4 on expression of PTEN and activation of AKT was assessed by immunoblotting PTEN AKT or phospho-AKT in the cell lysates with their antibodies respectively. c, Immunohistochemical (IHC) staining of 63 human lung adenocarcinoma tumors with anti-NEDD4 and anti-PTEN antibodies. The positive tumor samples were assessed and counted under microscope and listed in the table