Fig. 4.

Y288C protein activates signaling pathways in distinct subcellular compartments. a Phosphorylation of the Y288C receptor at the ER and Golgi was analyzed by western blot of the OptiPrep gradient ultracentrifugation resolved membrane fractions. MCF10A cells stably expressing PDGFRA WT or Y288C were serum-starved for 16 h and treated with or without monensin (500 nM) or BFA (1 μM) for 2 or 4 h respectively. b Cells were fixed and stained with anti-PDGFRA (green), anti-GM130 (Golgi marker; red) or anti-calnexin (ER marker; red), and DAPI (nucleus; blue) to examine the subcellular localization of PDGFRA after treatments. Bar, 25 μm. c Protein was extracted and analyzed for the activities of Akt, STAT3, STAT5, ERK1/2 signaling pathways by western blot with indicated antibodies. Corresponding total proteins were used as loading controls