Fig. 6

EGF treatment increases LAT3 expression on the plasma membrane. a and b, LAT3 expression levels were examined 5, 15 or 30 min after EGF stimulation of LNCaP (a) or PC-3 cells (b). c and d, LAT3 and Akt expression levels were examined in the presence or absence of MK2206 in combination with EGF in LNCaP (c) or PC-3 cells (d). GAPDH was used as loading control. e LAT3 protein levels were examined in the absence or presence of EGF in LNCaP cells by cell surface protein isolation; Na, K-ATPase was used as loading control. GAPDH was used to confirm cell surface fraction purity. f Ubiquitin and LAT3 were examined after immunoprecipitation with isotype IgG or anti-LAT3 IgG in the presence or absence of MK2206 or MG132 with EGF stimulation in LNCaP cells. Input of cell lysates pre-immunoprecipitation was examined using LAT3 and GAPDH antibodies. Ratio of IB: ubiquitin expression level is shown relative to Input: LAT3. All western blot images are representative of three independent experiments