Figure 2
CK2 activity in γδ thymocytes is modulated by TCR stimulation and activates AKT signaling. (a) In vitro CK2α activity in sorted γδ and αβ thymocytes (2 × 106 cells per sample) after 6 h of stimulation with anti-CD3 antibodies (CD3), plus soluble CD27-ligand (CD3+CD27) or plus 5 μM CX-4945 (CD3+CD27+CX); values were normalized to unstimulated control (dashed line). (b) Western blot analysis of (phospho)proteins implicated in AKT signaling, in γδ and αβ thymocytes (1 × 106 cells per sample) stimulated as in (a). (c) Proliferation (CFSE dilution assay) of γδ thymocytes after 7 days in culture with recombinant human IL-2 (rhIL-2) under the indicated conditions: medium only (Ctrl); anti-CD3 antibody stimulation (CD3); soluble CD27-ligand (CD27); their combination (CD3+CD27); and with 5 μM CX-4945 (CD3+CD27+CX). (d) Survival (% of live cells) of γδ thymocytes after 7 days of stimulation (or not, Ctrl for control) with anti-CD3 antibodies (CD3), plus soluble CD27-ligand (CD3+CD27), plus 5 μM of CX-4945 or 10 μM of MK-2206. Data in this figure are representative of at least three independent experiments; *P<0.05, ***P<0.001 (T-test).
