Supplemental Material 1


Supplemental Material 1: Disruption of Zap-70 from Jurkat cells and phosphorylation of Erk1/2.  A)  Zap-70 expression in Jurkat T cells.   Protein lysates from P116 (Zap-70 null) and P116.c139 (Zap-70 reconstituted) were separated by SDS-Page and immunoblotted with Zap-70 specific antibodies.  B)  Erk1 and Erk2 phosphorylation in OKT3/OKT4 stimulated Jurkat T cells used in phosphoproteomic analysis.  P116 (Zap-70 null) and P116.c139 (Zap-70 reconstituted) cells were stimulated with anti-CD3 (OKT3) and anti-CD4 (OKT4) antibodies.  Protein lysates were then separated by SDS-Page and immunoblotted with phospho-p44/p42 MAPK (Erk1 and Erk2) specific antibodies.

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