Data Supplement

Files in this Data Supplement:

• Supplemental Figure 1 (PDF, 165 Kb)
  - Flow cytometric analysis of thymocytes from chimeric mice used for imaging. (A) Representative data showing expression of CD4 and CD8 on GFP gated cells from P14-> CD11cYFP chimera (sample from Figure 4A). (B) Representative data of thymocyte populations from wt, CFP + P14, GFP -> CD11cYFP co-chimera (sample from Figure 4D). Ly5.1 was used here as a gating marker for the wild type donor population and GFP was used to gate the P14 donor population.
• Supplemental Figure 2 (PDF, 137 Kb)
  - Expression of CD5 and CCR7 in gated CD4+CD8+ thymocytes from the indicated mice. (A) Representative histogram showing expression of CD5 on CD4+CD8+ thymocytes from wild type (blue), TCRalpha-/- (red), and P14 Rag2-/- (green) mice. (B) Representative flow data showing CD5 and CC7 expression in the CD4+CD8+ thymocyte populations from wild type, CCR7-/-, TCRalpha-/-, and P14 Rag2-/- mice.
• Supplemental Video 1 (MOV, 669 Kb)
  - A 3D rotation showing YFP-labeled DCs (green) that are closely associated with cortical vasculature which is labeled with lectin-Texas Red (red). Second harmonic signal generated by collagen fibers of the capsule are shown in dark blue. Dimensions: 135x68x188 microns³. Corresponds to Figure 2C.
• Supplemental Video 2 (MOV, 755 Kb)
  - Example of a rare, motile host DC (red). Also a representative movie from a P14->CD11cYFP chimera with thymocytes in green. Migration path is time coded from blue to yellow. Dimensions: 153x160x34 microns³ x 24 minutes. Corresponds to Figure 3B.
• Supplemental Video 3 (MOV, 594 Kb)
  - A 3D rotation of a representative volume from a wild type->CD11cYFP chimera with thymocytes in green and CD11cYFP+ cells in red. Dimensions: 164x164x40 microns³.
• Supplemental Video 4 (MOV, 568 Kb)
  - A representative run from a wild type->CD11cYFP chimera showing thymocytes that migrate slower and have fewer contacts with DCs as compared P14->CD11cYFP (Video 5). Dimensions: 164x164x40 microns³ x 25 minutes. Corresponds to Figure 4A, top panel.
• Supplemental Video 5 (MOV, 789 Kb)
  - A representative run from a P14->CD11cYFP chimera showing thymocytes migrating at intermediate speeds and contacting DCs. Dimensions: 164x164x40 microns³ x 25 minutes. Corresponds to Figure 4A, bottom panel.
• Supplemental Video 6 (MOV, 156 Kb)
  - An example of a thymocyte (green) making a sharp turn after a brief encounter with a DC (red) from a wild type->CD11cYFP chimera. Dimensions: 71x96x35 microns³x 24 minutes.
• Supplemental Video 7 (MOV, 343 Kb)
  - An example of a single thymocyte (green) making successive encounters with multiple DCs (red) from a P14->CD11cYFP chimera. Dimensions: 109.5x138x41 microns³ x 25 minutes.
• Supplemental Video 8 (MOV, 223 Kb)
  - An example of a thymocyte (green) making contacts with the cell body of a DC (red) from a wild type->CD11cYFP chimera for the duration of the run. Dimensions: 164x51x41 microns³ x 18.5 minutes.
• Supplemental Video 9 (MOV, 118 Kb)
  - An example of a thymocyte (green) contacting the tips of a DC (red) from a P14->CD11cYFP chimera.
• Supplemental Video 10 (MOV, 895 Kb)
  - An example of a thymocyte (blue) enclosed by a DC (yellow) from a wt, CFP (blue) and P14, GFP (green)->CD11cYFP (yellow) high-density chimera. A single z slice over time is shown.
• Supplemental Video 11 (MOV, 2.51 Mb)
  - A representative run from a wt, CFP (blue) and P14, GFP (green) -> CD11cYFP co-chimera. Corresponds to Figure 6. Dimensions: 173x142x80 microns³x15 minutes.
• Supplemental Video 12 (MOV, 854 Kb)
  - Rotation of an imaging volume showing DC (orange) and blood vessels (red). The tracks of 5 thymocytes that migrate >13 microns/min and move toward the medulla are highlighted. The signal from thymocytes is not shown.