Fig. 7. A_2AR suppresses the cytoskeletal polarization of CTL.

a Percentage of in vitro CL4 CTL with a uropod ± 1 µM CGS-21680 ± SEM. N = 4 independent experiments, 220/648 T cells analyzed. Representative images of CL4 T cells (labeled with ‘T’) in a field with Renca APC treated ± 1 µM CGS-21680. Arrows indicate uropods. Scale bar = 5 µm. b Frequency of CL4 T cells that form a tight cell couple upon contact with a Renca APC incubated with 2 µg/ml K^dHA peptide ± SEM. N = 2 independent experiments, 68/234 T cells analyzed. c, d Imaging of the interaction between in vitro CL4 CTL with K^dHA-pulsed Renca tumor cell targets treated with NECA ±1.25 μM A2aR antagonist ZM 241385. c Percentage of cell couples with translocation (1-way ANOVA) ±SEM. N = 28-132 cell couples per condition over ≥2 experiments. Alternate analysis of pooled data by proportion’s z-test yields p < 0.001 for all comparisons between NECA only-treated and control or ZM 241385-treated samples. d Time until the formation of first off-interface lamella (Kaplan–Meier survival analysis (Log Rank)). P < 0.01 all comparisons. e In vitro CL4 CTL interacted with RencaHA cells incubated with 2 µg/ml K^dHA peptide ± 1 µM CGS-21680. The ratio of Fura-2 emissions at 510 nm upon excitation at 340 nm over 380 nm is given relative to time of tight cell coupling as the mean ± SEM. N = 2 independent experiments, 13/42 T cells analyzed. f CL4 T cells were primed in vitro using anti-CD3/CD28 mAb. NECA ± 1.25 µM ZM 241385 were added at 0 h and ³H-thymidine for the last 8 h of cell culture. Proliferation was quantified by ³H-thymidine incorporation (cpm) and is given as the mean ± SEM (N = 3 experiments). Source data are provided in Supplementary Data 6 (all but the calcium data) and Supplementary Data 7 (calcium data).