Figure 5. Epidermal Flot1^-/- CD8⁺ T_RM migration.

A. Experimental layout for the generation of epidermal T_RM with HSV_TOM-OVA infection. B. Ratio of OT-I and Flot1^-/- OT-I T cells recovered at day 7 from HSV_TOM-OVA infected skin. C. 2PM image sequence of memory OT-I and Flot1^-/- OT-I T cells showing both xy and xz views of epidermis over 2 h of continuous imaging. OT-I cells and cell tracks in white, and Flot1^-/- OT-I cells and tracks in green. Dermal collagen visualized by second harmonic generation shown in blue. Scale bar 50 μm; Time in h:min:s. D. Representative time-coded outlines of migrating memory OT-I and Flot1^-/- OT-I T cells in the epidermis. Scale bar, 20 μm. E. Shape factor of epidermal OT-I and Flot1^-/- OT-I T cells. Pooled from one experiment with n = 2 (OT-I) or n = 3 (Flot1^-/- OT-I) mice. F-H. Speed (F), arrest coefficient (G) and meandering index (H) of epidermal OT-I or Flot1^-/- OT-I T cells. Red lines depict median, each dot represents the average value for an individual track. I. Mean displacement versus time for datasets in E-G. Numbers indicate motility coefficient in μm²/min. Data in B are pooled from 2 independent experiments (n = 5) and data E to I were pooled from 3 independent experiments (n = 10). Data in F were analyzed using unpaired Student’s t-test, data in E, G and H using Mann-Whitney test. ***, p < 0.001.