Extended Data Fig. 2: The traction force exerted by T cells is enhanced by PIEZO1 blockade.

a, The traction force of CD8⁺ T cells on the polyacrylamide (PAA) gels with different stiffness, ranging from 0.3 kPa to 10.0 kPa. Scale bar, 5 µm. n = 60. b, The traction force of naive and activated CD8⁺ T cells from murine spleens (n = 111 for naive; n = 102 for active). Scale bar, 5 µm. c-g, Activated CD8⁺ T cells from murine spleens (c, d) or healthy donors (e, f, and g) were transfected with shRNAs targeting PIEZO1. The GFP⁺ T cells were sorted and the knockdown efficiency of PIEZO1 was confirmed by real-time PCR (c, e) or flow cytometry (d, f). In some experiments, sorted GFP⁺ T cells were performed the TFM (g, n = 100). Scale bar, 5 µm. h, Activated OT-1 CTLs were treated with fluid shear stress, squeezing force, hypertonic or hypotonic osmotic buffer. The traction force of CTLs was determined by TFM (n = 60). Scale bar, 5 µm. i, The traction force of activated OT-1 CTLs pretreated with Veh., cytochalasin D (1 µM) (n = 95). Scale bar, 5 µm. j, OT-1 CTLs were treated with cytochalasin D (1 µM), and then co-cultured with OVA-B16 cells. CD45⁻ tumor cells apoptosis was determined by flow cytometry. Mann-Whitney test (b and i), two-tailed Student’s t-test (j) or one-way ANOVA followed by Bonferroni’s test (a, c-h). The data represent mean ± s.d.

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