Figure 5
From: Nck1 depletion induces activation of the PI3K/Akt pathway by attenuating PTP1B protein expression
Nck and PTP1B interaction. (A) HEK293 cells were transfected with FLAG-PTP1B and pRK5 (empty vector) or HA-Nck1. Total cell lysates were subjected to anti-HA immunoprecipitation followed by immunoblot using indicated antibodies. Data are representative of three independent experiments showing similar results. (B) HEK293 cells were transfected with FLAG-PTP1B and pcDNA3.1 (empty vector). Total cell lysates were subjected to anti-FLAG immunoprecipitation followed by immunoblot using indicated antibodies. (C) Total cell lysates from HEK293 or HepG2 cells were subjected to IgG or Nck immunoprecipitation followed by immunoblot with indicated antibodies. The vertical line in the immunoblot is to indicate that samples are separated by empty lanes. (D) Cell lysates from HEK293 cells overexpressing FLAG-PTP1B were incubated with GST, GST-Nck1, GST-SH2 domain of Nck1, GST-SH3 domains of Nck1 for 3 h followed by immunoblot with anti-FLAG antibody. Ponceau staining reveals the GST-fusion proteins. Data are representative of three independent experiments showing similar results.