Myo9 enables T cell-mediated global skin immunity. (A) 2PM images of WT (closed arrowheads) and Myo9b^−/− (open arrowheads) OT-I T_RM at indicated time after OVA_257–264 peptide tattooing. Bars, 20 µm. (B and C) Quantification of 2PM data pooled from six image sequences in two mice from one experiment. WT and Myo9b^−/− OT-I T_RM speeds (B) and sphericity (C) at indicated time after OVA_257–264 peptide tattooing. (D) In vitro killing efficacy of sorted WT and Myo9b^−/− OT-I T_RM. Pooled from two independent experiments. (E) Experimental strategy to generate WT and Myo9b^−/− T cells in skins of Prf1^−/− mice, followed by HSV-1_TOM-OVA challenge. (F) Viral titers at 60 h after infection with HSV-1_TOM-OVA after transfer of WT or Myo9b^−/− OT-I T cells in OVA_257–263–tattooed and untreated contralateral skin flank. Each dot represents skin viral titers from one mouse. Pooled from two experiments. The dotted line corresponds to viral titers after “no cell” transfer (i.e., untreated Prf1^−/− mice). Statistical analysis: ANOVA with Tukey’s multiple comparison test (B and C); ANOVA (D); Kruskal Wallis test against “no cells” with Dunn’s multiple comparison test (F). *, P < 0.05; **, P < 0.01; ***, P < 0.001. Horizontal bars depict mean.