Figure 4
From: Proteomic analysis of tyrosine phosphorylation during human liver transplantation
Nck-1 expression and subcellular localization in human liver upon I/R. A. Immunoblot analysis of Nck-1 from total fractions of 0, 10 and 60 min ischemia (I0, I10, I60) and 0, 10 and 60 min reperfusion (R0, R10, R60) liver protein extracts (WCL: Whole Cell Lysate) normalized to Intersectin (Int). N = 2 on 3 independent pools of 3 liver biopsy protein extracts. B. Immunoblot analysis of Nck-1 from insoluble (P100 – lower panel) and soluble (S100 – upper panel) fractions obtained after centrifugation of liver biopsies homogenized in the presence of 150 mM KCl respectively normalized to Ribophorin (Rib) and Intersectin (Int) for the 3 ischemia time points – 0 (I0), 10 (I10) and 60 (I60) min and the 3 reperfusion time points 0 (R0), 10 (R10) and 60 (R60) min. N = 2 on 3 independent pools of 3 liver biopsy protein extracts. C. Immunohistochemical detection of and Nck-1 on 8 μm liver tissue sections 0 and 60 min post-cold ischemia (I0 and I60) and 60 min post-reperfusion (R60). Cells were counterstained with haematoxylin (cells nuclei appear in blue). A representative experiment out of the 6 performed on independent ischemic and reperfused livers is shown. Scale bar = 10 μm.