Fig. 2: TCRs are released through the F-actin/ezrin-enriched microvillar protrusions of activated T cells on iAb. | Nature Communications

Fig. 2: TCRs are released through the F-actin/ezrin-enriched microvillar protrusions of activated T cells on iAb.

From: Trogocytic molting of T cell microvilli upregulates T cell receptor surface expression and promotes clonal expansion

Fig. 2

a CD4+ T blasts expressing TCRζ_GFP were stained with either CTV or CTB (cholera toxin B) and stimulated with iAb (10 μg/mL) or sAb (10 μg/mL) for 3 h. Localization of TCRζ_GFP in resting (PLL, top) or activated conditions (TCRζ_GFP or CTB, bottom) was observed. Yellow arrows represent internalized or released TCRζ_GFP at each condition. TCR ζ intensity (sum of 3D MFI) was presented as inside intensity divided by total intensity. For each single experiment, 50 cells were randomly selected and analyzed. Results are representative of three independent experiments ± SD. Statistics was performed using one-way ANOVA with post hoc Tukey’s multiple comparisons test vs. PLL. Source data are provided as a Source Data file. b CTV and anti-TCRβ-Alexa488-pre-stained naive CD3+ T cells were stimulated as in (a), fixed, permeabilized, and stained with either anti-ezrin antibody or phalloidin-TRITC. TCRβ localization across the microvilli in boxed area (cyan) was represented with pseudo-color coding according to fluorescence intensity. Yellow arrows represented colocalization of TCRβ with either ezrin or F-actin in resting condition. Red and yellow arrow represented dissociation of TCRβ from ezrin and F-actin in sAb condition. All images were acquired by AiryScan confocal microscopy and reconstructed for the 3D by using Zeiss ZEN software. These results were independently repeated three times on more than 10 randomly selected cells each time.

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