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Figure 2


Fig. 2. GDNF stimulates interaction of RET with SH2-Bß in PC12-GFR{alpha}1-RET cells. (A) PC12-GFR{alpha}1-RET cells were transfected with pcDNA3, pcDNA3-myc-SH2-Bß, or pcDNA3-myc-R555E respectively. Cells were treated or not with 100 ng/ml GDNF for 10 minutes before lysed on ice and collected. Then, cell lysates were immunoprecipitated (IP) with anti-myc or anti-RET antibodies, followed by immunoblotting (IB) with anti-RET or anti-myc antibodies. Cell lysates were also subjected to immunoblotting with anti-RET or anti-myc antibodies. (B) Cultured PC12 cells or PC12-GFR{alpha}1-RET cells were treated or not with 100 ng/ml GDNF for 10 minutes before they were lysed and collected. Cell lysates were immunoprecipitated with anti-RET or anti-SH2-Bß antibodies, followed by immunoblotting with anti-SH2-Bß or anti-RET antibodies. Expression of RET and SH2-Bß in cells was determined by immunoblotting with anti-RET or anti-SH2-Bß antibodies. IP, immunoprecipitation; IB, immunoblot.