Fig. 4: PEG Regulatory T cells do not disrupt prolonged macrophage:PEG Th1 contacts.

A Control Th1 cells or Tregs and PEG Th1 cells were stained with cell tracker dyes (CMFDA; green, and CMTMR; red, respectively) and co-cultured with stained uninfected or L. major- infected macrophages (CMAC; blue). B A time series micrograph of co-cultures of macrophages, control and PEG Th1 cells in the absence or presence of L. major infection at indicated times. Time stamps in min:sec represent elapsed time of the recordings. C Real-time contact duration between control (blue dots) or PEG (red dots) Th1 cells and macrophages in the absence or presence of L. major infection. Each dot represents a single contact. Black lines represent median values. Statistical analysis: Mann–Whitney U-test, ns not significant. Data combined from three independent experiments. D Mean 3D track velocity of control (blue dots) or PEG (red dots) Th1 cells cultured with macrophages in the absence or presence of L. major infection. Each dot represents a single cell track. Black lines represent median values. Statistical analysis: Mann–Whitney U-test, ns not significant. Data combined from three independent experiments. E A time series micrograph of co-cultures of L. major-infected macrophages, PEG Th1 cells in the presence of control or PEG Tregs at indicated times. Time stamps in min:sec represent elapsed time of the recordings. Real-time contact duration between control or PEG Tregs and macrophages (F) and mean 3D track velocity (G), each dot representing a single contact. Black lines represent median values. Statistical analysis: Mann–Whitney U-test. Data combined from three independent experiments. Source data are provided as a Source Data file. H Real-time contact duration between PEG (red dots) Th1 cells and L. major-infected macrophages. Each dot represents a single contact. Black lines represent median values. Statistical analysis: Mann–Whitney U-test, ns not significant. Data combined from three independent experiments. I Mean 3D track velocity of PEG (red dots) Th1 cells. Each dot represents a single cell track. Black lines represent median values. Statistical analysis: Mann–Whitney U-test, ns not significant. Data combined from three independent experiments. Track velocities of control (blue) or PEG (red) Th1 cells cultured with uninfected or L. major-infected macrophages plotted against confinement ratio (J) and relative percentage of confined slow and meandering fast tracks (K). Data combined from three independent experiments. Track velocities of PEG (red) Th1 cells cultured with L. major-infected macrophages in the presence of control or PEG Tregs plotted against confinement ratio (L) and relative percentage of confined slow and meandering fast tracks (M). Data combined from three independent experiments. N Flow cytometry analysis of L. major-infected macrophage surface molecules. Representative data from three independent experiments. Source data are provided as a Source Data file.