Fig. 2: Essential roles of PD-1 and CD80 for T cell TEM.

a In vitro migration assay of iTregs treated with anti-mouse PD-1 (Rmp1-14), anti-mouse PD-L1 (10 F.9G2), or rat IgG (2A3), loaded in Boyden chambers with or without LEC. b Same as in a, in vitro migration of blocking mAb-treated Teffs. c Comparison of blocking intact mAb (2 μg/mL) and F(ab)’2 mAb (1 μg/mL) treatment of iTregs for TEM. d WT, PD-1-deficient (PD-1^−/−), and PD-L1-deficient (PD-L1^−/−) iTregs TEM. e Time-lapse imaging of WT, PD-1^−/−, PD-L1^−/− iTreg movements during TEM. f In vitro migration of human Tregs or Teffs blocked with 5 μg/mL anti-human PD-1 (EH12.2H7), anti-human PD-L1 (29E.2A3), or mouse IgG1. g iTregs as in a or Teffs as in b treated with 2 μg/mL anti-mouse PD-1 (Rmp1-14), anti-mouse CD80 (1G1), or rat IgG (2A3). h Teffs as in b treat with various doses of anti-CD80 (1G1) or anti-PD-1 (Rmp1-14). i iTregs as in a loaded in Boyden chambers coated with 1 μg/mL mouse PD-L1 Ig, CD80 Ig, LTβR Ig, or mouse IgG1. j, k Immunoblotting for Akt, NFκB-p65, and ERK phosphorylation after PD-L1 ligation of iTregs (j) and Teffs (k) stimulated for the indicated times with 1 μg/mL mouse PD-L1 Ig, CD80 Ig, or IgG1 coated on wells. Relative band intensities shown. Data representative of 3 independent experiments (a–k). Mean ± SEM. *p < 0.01 by one-way ANOVA with Sidak’s multiple comparisons test in the same group (a–d, f–i), p < 0.0001 by unpaired t-test (e), or p < 0.001 by unpaired, two-tailed t-test with Welch’s correction (j, k). Source data are provided as a Source Data file.