Legends to supplementary figures
Figure S-1
FACS sorted PP DC generate high CD8 effector cells. P14xT-GFP cells were cultured with gp33-41pulsed DC from different lymphoid organs that were either isolated by negative selection (85-90%
CD11c+) or by negative selection plus FACS sorting (>98% CD11c+).  expression (% h
cells and mean fluorescence intensity [MFI]) was compared after 6 days. (a) FACS plots from a
representative experiment. DC from PP, PLN, and spleen isolated by negative selection (left) or
negative selection plus FACS-sorting (right) and the corresponding expression on the resulting
effector T cells. Smaller FACS plots show staining of T cells with isotype-matched control MAb. In
FACS plots showing DC, the frequency of CD11c+ Linneg cells is shown. FACS plots showing T cells
provide the frequency of high cells and, in parenthesis, the MFI for  of the entire T cell
population. (b) MFI for 4ß7 was not significantly different between T cells activated with PP DC
isolated by negative selection vs. PP DC isolated by negative selection plus FACS-sorting (n = 6-8
experiments/group; *** p<0.001 vs. corresponding CD8PP-DC). (c) Only PP DC, but not the sorted
CD11cneg fraction from PP, induced 4ß7 on anti-CD3/CD28 activated P14xT-GFP cells (n=3
experiments/group; ** p<0.01 vs. anti-CD3/CD28 alone). Data are presented as mean ± SEM and were
analyzed by one-way ANOVA with Bonferroni correction.
Figure S-2
PP DC generate high cells with comparable efficiency from total T cells isolated from P14xT-GFP
splenocytes and from pre-sorted GFP+/neg T cells. Negatively isolated T cells or sorted
4ß7neg/GFP+ T cells were cultured for 6 days with gp33-41-pulsed PP DC or PLN DC and then analyzed
for expression (small plots show staining with isotype control MAb vs. GFP). Representative
FACS plots show the starting P14xT-GFP populations isolated by negative selection alone (top left;
square indicates gate setting for sorting), and after FACS-sorting (bottom left). On the right,
expression is shown on effector populations after activation with peptide-pulsed PP DC or PLN
DC. FACS plots provide the frequency of high cells and, in parenthesis, the MFI for  of the
entire T cell population (small plots show staining with isotype control MAb vs. GFP).
Figure S-3
Expression of on specific peptide-MHC-I tetramer binding CD8+ effector T cells. P14xT-GFP
cells were cultured for 6 days with gp33-41-pulsed DC isolated from PP, PLN, or spleen, and then
analyzed for expression and gp33-41-H-2Db tetramer binding. Small FACS plots show staining
with a control MHC tetramer (np396-404-H-2Db) and with isotype-matched control MAb for anti-4ß7.
The figure shows one representative experiment of five performed with similar results.
Figure S-4
Stimulation by FACS sorted DC induces CD8PP-DC that respond more efficiently to TECK/CCL25 as
compared to CD8PLN-DC or CD8Sp-DC. P14xT-GFP cells were cultured for 6-7 days with gp33-41-pulsed
DC isolated from different lymphoid tissues and compared for their ability to migrate to TECK in
Transwell chemotaxis assays. TECK mediated chemotaxis of P14xT-GFP cells that were cultured in
parallel with unsorted negatively selected PP DC is shown for comparison. The figure shows one
experiment of two performed with similar results.
Figure S-5
PP DC generate a gut homing phenotype in CD8+ T cells from OT-IxRAG-/- mice. OT-IxRAG-/- CD8+
cells were cocultured for 6 days with antigenic peptide-pulsed PP DC or PLN DC and then compared for
(a)  expression (n = 3, * p<0.05 by two-tailed paired Student’s t test) and (b) chemotaxis to TECK
(n = 2 experiments for 300 nM TECK (left), and 1 dose/response determination of TECK ranging from
100 to 500 nM (right). (c) Homing properties of OT-IxRAG-/- T cells activated with either PP DC or
PLN DC (n = 1 experiment).
Figure S-6
The preferential homing of CD8PP-DC to the small intestine lamina propria (LP) is partially blocked by
anti-TECK MAb. Naïve CD8+ P14xT-GFP cells were cultured with gp33-41-pulsed PP DC or PLN DC.
After 6 days, effector T cells were used for competitive homing experiments in mice treated with either
anti-TECK MAb or isotype matched control MAb. (a) FACS plots from 2 independent experiments
show migration of CD8PP-DC and CD8PLN-DC to the small intestine. The same total number of events is
shown in each plot. Note the reduced number of homed CD8PP-DC in gate R3 after anti-TECK treatment.
The homing index (HI) was determined as described in Methods. (b) Anti-TECK selectively reduced
homing of CD8PP-DC to the small intestine, but not to other tissues. Error bars represent range of 2
experiments.