Figure Supplementary 5: The stability of pre-existing skin T_RM cell populations after viral challenge.

a Experimental schematic related to Fig. 5a-c. Mice received Thy1.1⁺ gBT-I T_EFF cells and were treated with DNFB. After >30d Thy1.1⁺ gBT-I T_CIRCM cells were depleted using Thy1.1 antibody. A separate cohort received CD45.1⁺ gBT-I T_N cells and was infected with HSV. >30 d post-infection, splenic CD45.1⁺ T_CIRCM cells from these donors were transferred into mice containing DNFB-lodged Thy1.1⁺ gBT-I T_RM cells and these recipients were then infected with HSV above DNFB-treated skin, which was analysed >30 d post-infection. b Experimental schematic related to Fig. 5d-f. Mice received Thy1.1⁺ P14 T_EFF cells and were treated with DNFB. After >30d Thy1.1⁺ P14 T_CIRCM cells were depleted using Thy1.1 antibody. >7 d later mice were infected with HSV above DNFB-treated skin and transferred CD45.1⁺ OT-I T_EFF cells. DNFB-treated skin was analyzed >30 d post-HSV infection. c Experimental schematic related to Fig. 5g-i. Mice received Thy1.1⁺ gBT-I (T_N) cells and were infected with VV-gB on the lower skin flank e.c. >30 d post-infection Thy1.1⁺ gBT-I T_CIRCM cells were depleted using Thy1.1 antibody. >7 d later mice were challenged with HSV above VV-challenged skin and CD45.1⁺ OT-I T_EFF cells were transferred 2d later. VV-challenged skin was analysed >20 d post HSV-infection. d Experimental schematic related to Supplementary Fig. S5e. μMT mice received Thy1.1⁺ gBT-I (T_N) cells and were infected with HSV on the lower left flank. >25 d post-infection, mice were treated with Thy1.1 antibody to remove Thy1.1⁺ gBT-I T_CIRCM cells. 10d post-antibody treatment mice were challenged with HSV on the upper left flank and transferred CD45.1⁺ OT-I T_EFF cells i.v. 2d post-infection. e Number of Thy1.1⁺ gBT-I and CD45.1⁺ OT-I T_RM cells in HSV-challenged skin >20 d post-HSV infection. Data are pooled from 2 experiments with n = 7 or 8 mice per group. **p=0.0059, two-tailed Mann Whitney U-test. Bars represent the mean.