Figure 5.

ITK-mediated TCR/ligand induced signaling in the skin impairs the development of sIELs by promoting their apoptosis. A. Skin cell preparations of KN6 transgenic mice of wild type and ITK^−/− backgrounds were analyzed for transgenic Vγ2⁺ sIELs by flow cytometry. Percentages of the transgenic sIELs were indicated. One representative of three independent experiments is shown. B and C. Ear epidermal sheets of ITK-sufficient and knockout KN6 mice were stained and observed under a fluorescent microscope (Olympus BX61) for transgenic Vγ2⁺ sIELs (B), average numbers of which per field at the 200x amplification were plotted (C). Data were obtained from three experiments. ** P < 0.01. D. Lower percentages of apoptotic KN6 transgenic sIELs on ITK^−/− than wild type background. The percentages of apoptotic sIELs were calculated based on ratios of numbers of apoptotic vs. total sIELs from in situ TUNEL analyses of ear epidermal sheets, as shown in the Supplementary Fig. 3. Data shown were obtained from at least four mice of each genotype in two independent experiments. * P < 0.05. E. The development of KN6 transgenic sIELs in β2m^−/−TCRδ ^−/− recipients from adoptively transferred fetal thymic ITK-sufficient or knockout fetal thymic KN6 transgenic γδ T cells. Ear epidermal sheets of the recipients were analyzed for donor-derived sIELs by in situ immunofluorescent staining (Olympus BX61). Data presented is one representative from three mice of each genotype.