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Fig. 5 CRK and NCK family SH2 domains have overlapping neuronal binding partners. (A) Anti-phosphotyrosine (α-pY) and anti-tubulin western blots of extracts from Neuro2A cells treated with (+) or without (−) hydrogen peroxide (H2O2). Molecular weight markers in kilodaltons are indicated. (B) Representative coomassie-stained gels used for MS analysis of GST-NCK1-SH2 and GST-NCK2-SH2 pulldowns using extracts from A. *Indicates full-length GST-SH2 fusion (breakdown products are major bands below). (C) Previously [31], we found 46 proteins from Neuro2A cells were induced by H2O2 to bind to both the SH2 domains of CRK and CRKL. 29 of these were also found induced by H2O2 to interact with the SH2 domain of either NCK1 or NCK2. The pie chart indicates the proportions of the 29 proteins that interact with one or both NCK family member's SH2 domain. Binding partners involved in cell migration (see Fig. 6) are listed.