Th1 cells form FAs. (A–D) Representative migrating Th1 cells expressing Talin-EGFP in confinement on (A) ICAM-1, (B) FN, (C) PLL, and (D) PLL-PEG. (E) Representative image of a confined Th1 cell fixed and stained for endogenous talin. (F and G) Quantification of the number of FA-positive cells when expressing (F) Talin-EGFP or (G) vinculin-EGFP compared across surface treatments. (H) A comparison of FA lifetime between Th1 cells and fibroblasts expressing Vinculin eGFP (representative images on right). (I and J) Representative Th1 cells expressing (I) Talin-EGFP or (J) Integrin β₃-Emerald in confinement on a micropatterned substrate. The left portion of the field of view is FN coated and the right portion is passivated. Number of biological replicates (colored dots): (F) Passivated, 36 cells from three experiments; PLL, 53 cells from two experiments; FN, 140 cells from five experiments; ICAM-1, 40 cells from three experiments. (G) Passivated, 21 cells from two experiments; PLL, 38 cells from four experiments; FN, 94 cells from four experiments; ICAM-1 169 cells from three experiments. (H) Th1 n = 10; Fibroblast n = 10. Each replicate represents the average turnover of ≥5 FAs per cell. Statistical tests: (F–H) Two-tail unpaired parametric t test was used to compare the replicates.