Fig. 2: Shp2 is a protein that interacted with EphA2 in the NPC cells. | Cell Death & Disease

Fig. 2: Shp2 is a protein that interacted with EphA2 in the NPC cells.

From: Y772 phosphorylation of EphA2 is responsible for EphA2-dependent NPC nasopharyngeal carcinoma growth by Shp2/Erk-1/2 signaling pathway

Fig. 2

a IP-mass spectrometry (MS) analysis of the proteins interacted with EphA2. Total cell proteins from 5-8F NPC cells were subjected to co-immunoprecipitation with anti-EphA2 antibody. The coimmunoprecipitated protein complex was resolved on SDS-PAGE and Coomassie blue staining, and then the bands were retrieved and analyzed by MS analysis. b Identification of Shp2 as a protein that interacted with EphA2 by MS analysis. The amino acid sequence of a doubly charged peptide with m/z 396.2391 Da was identified as NAAEIESR and Mascot search showing the peptide matched with Shp2. c Co-IP confirming the interaction of Shp2 and EphA2. Total cell proteins from the 5-8F and CNE2 NPC cells (left) and HEK293 cells ectopically expressing EphA2 (right) were prepared and subjected to immunoprecipitation (IP) with anti-EphA2 antibody followed by immunoblotting with antibodies against Shp2 or EphA2. d Immunofluorescence showing the colocalization of EphA2 (green) and Shp2 (red) in the NPC cells. 5-8F and CNE2 cells were incubated with mouse anti-EphA2 and rabbit anti-Shp2 antibodies followed by staining with DyLight® 488 anti-mouse IgG and DyLight® 594 anti-rabbit IgG, and observed by confocal fluorescence microscopy. Scale bar = 10 μm. e Co-IP showing that EphA2-Y772A mutation does not disturb the interaction of EphA2 and Shp2 in the NPC cells. Total cell proteins from the 5-8F and CNE2 cells expressing EphA2-WT or EphA2-YA were prepared and subjected to immunoprecipitation (IP) with anti-EphA2 antibody followed by immunoblotting with antibodies against Shp2 or EphA2.

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