Nck-1 tyrosine phosphorylation status upon I/R. A. Nck-1 sequence: peptides identified by mass spectrometry are represented underlined, predicted tyrosine phosphorylation site are in red bold. B. Representative MS/MS spectrum corresponding to an Nck-1 specific peptide. C. Prediction of tyrosine-phosphorylation sites using the NetPhos software. D. Immunoblot analysis of Nck-1, using anti-Nck-1 antibody (Ib Nck-1) and Nck-1 tyrosine-phosphorylation status, using anti-phosphotyrosine antibody (Ib PY) following Nck-1 immunoprecipitation (Ip) on 60 min ischemia (I60) and 60 min reperfusion (R60) fractions. The band corresponding to Nck-1 is indicated by a black arrow. Immunoglobulins Heavy Chains (Ig HC) are indicated by a purple arrow. N = 2 on 3 independent pools of 3 liver biopsy protein extracts.