Fig. 5: Differential crosstalk between migrating iTregs or Teffs and LECs.
From: PD-L1 signaling selectively regulates T cell lymphatic transendothelial migration
a Flow cytometry analysis of PD-1 on iTregs or Teffs after 3 h of TEM across mouse LECs. Values are the ΔMFIs for the migrated and non-migrated populations. b, c Flow cytometry analysis of PD-1 on human Tregs or Teffs (b) or PD-L1 on mouse or human LECs (c) after 3 h of TEM across LECs. d, e Analysis of interaction of PD-L1 with PD-1 (d), or with CD80 (e) on the interface (arrowhead) of CFSE-labeled iTregs or Teffs and LECs after 3 h of TEM. Colocalization coefficient and Pearson’s correlation of T cell PD-1 or CD80 interaction with LEC PD-L1 shown. Each dot represents the region of interest (ROI) of the interface. Percentage of CD80 at the T-cell-WT or PD-L1−/−LEC interface also shown. f–i Analysis of CD80 Ig binding to PD-L1 on LEC by immunohistochemistry (f, g), flow cytometry (h), and immunoprecipitation (i). j Teffs bind immobilized PD-L1 Ig (left panel) which promote TEM (right panel). k Analysis of NFκB-p65 activation in LEC 30 min after incubation with iTregs or Teffs. Arrows indicate the nuclear translocated NFκB-p65. Magnification ×60 (d–g, k), scale bar 5 μm (d), 7 μm (e, k), and 14 μm (f, g). Data representative of 3 independent experiments. Mean ± SEM (a–c, h, j). *p < 0.01 by one-way ANOVA with Sidak’s multiple comparisons test (c, j) and unpaired, two-tailed t-test with Welch’s correction (a, b, h). Source data are provided as a Source Data file.
