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. Author manuscript; available in PMC: 2009 Oct 22.
Published in final edited form as: J Immunol. 2009 Apr 1;182(7):4093–4106. doi: 10.4049/jimmunol.0803317

Figure 3.

Figure 3

The human Lyp1 R620W (T1858) polymorphism functions as a hypomorph in the context of Csk. (A) Csk and wild type or R620W Lyp1 were misexpressed in Jurkat cells together with GFP. Upper panel represents whole cell lysates of transfected cells probed for either myc-tagged Lyp1 or Csk. (B) Histograms correspond to samples in (A) and represent intracellular phospho-Erk staining of transfected Jurkat cells stimulated through the TCR by C305 or with PMA. GFP high, intermediate and low gating is shown. Gray histogram represent unstimulated transfectants for comparison. Black histograms represent stimulated transfectants as identified in (A). (C) Bar graph corresponding to samples generated and stimulated as in (A) and (B). % phospho-Erk positive cells are defined as in Figure 2(C). Error bars represent SEM. (D) Intracellular calcium levels of Lyp1/Csk transfected Jurkat cells (gated for co-transfected CD16). Fluo3/Fura Red ratio was monitored by flow cytometry before and after C305 stimulation of fluorophor-loaded transfectants. Solid line represents Csk+ Lyp1 R620*C1858 wild type allele. Dotted line represents Csk+ Lyp1 W620*T1858 risk allele. Plots, western, and calcium flux are representative of at least three independent experiments.