Patchy depletion and recovery of ear skin γδ T cells. (A) Frequencies of γδTCRhi epidermal (left) and γδTCRlo dermal (right) γδ T cells in nondepleted (ctrl.) and 7-wk-ago–depleted (7w) Tcrd-GDL mice. Pooled data from two independent experiments, one dot represents one mouse, mean. (B) Longitudinal in vivo two-photon imaging of Tcrd-GDL ear skin. Images were acquired from the same mouse at indicated time points after γδ T cell depletion. As reference, a mouse was injected with PBS (ctrl.). Green: γδ T cell eGFP by Tcrd-GDL mice, shown are representative images of two independent experiments. Bars, 50 µm (see Video 3). (C) Epidermal sheets of DTx- and PBS (ctrl.)–treated Tcrd-GDL mice 4 wk after γδ T cell depletion. Representative fluorescence microscopy images out of two independent experiments with n = 1–2 mice each; blue: Vγ5; red: CD3; white: DAPI nuclear staining. Bars, 50 µm. (D) Epidermal sheets of hetero- and homozygous Tcrd-GDL mice were stained with anti-CD3ε (red) 18 d after depletion (DTx) or PBS injection (ctrl.). Bars, 100 µm. Staining was validated in three experiments with n = 1–2 mice each.
