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. 2017 Jul 7;8:789. doi: 10.3389/fimmu.2017.00789

Figure 3.

Figure 3

ITAM and SYK-dependent, but Src-independent SLP-76 phosphorylation. S2-cells were co-transfected with the indicated proteins. Equal numbers of FACS-enriched GFP+ cells were analyzed by western blot. (A) Analysis of SLP-76 and CD3ζ phosphorylation upon co-expression with SYK, ZAP70, Fyn, or Csk. (B) S2-cells were co-transfected with GFP, SLP-76, CD3ζ, and SYK. SLP-76 phosphorylation was analyzed in GFP+ cells upon treatment with the SYK-family kinase inhibitor SKY IV, the Src-family kinase inhibitor PP1 or DMSO as a carrier control. (C) Analysis of SLP-76 phosphorylation upon co-expression of GFP, SLP-76, SYK, and either wild-type (wt) CD3ζ or the indicated CD3ζ ITAM-mutants. (D) Analysis of SLP-76 phosphorylation upon co-expression of GFP, SLP-76, CD3ζ and either wild-type (wt) SYK or the indicated SYK SH2 domain mutants. All western blots are representative of at least three independent experiments.