Supplementary Information accompanies the paper on Cellular ...
Day 5
Day 0
2hrs
RESEARCH ARTICLE IP
Isolate T cells from tissues
Stain for KLRG-1 ex vivo
Isolate T cells from tissues
KLRG-1 stained in vivo
IV
LCMVArm
PBS 2hrs
IP
IV
LCMVArm
KLRG-1 APC
in vivo 500
ex vivo 400
245
7643
400
233
4747
300
Cellular & Molecular Immunology advance online publication, ; doi:10.1038/cmi.2014.48 300 SPL
200
200
Supplementary Information accompanies100 the paper on Cellular & Molecular Immunology’s website 0(http://www.na49.6 50.4 0 10 10 10 ture.com/cmi). 5064 250 365 Supplementary Figure 1 3
4
Supplementary Figure 2
100
5
0
58.5 0
400
41.5 103
104
354
105
4317
200 300
iLN
150 200
100
100
50 0
0 1500
33.7 33.7
66.3 103
104
318
105
7534
0
0 1000
35.4 35.4
64.6 103
104
340
105
5709
800
1000
600
LVR
400
500
200
0
300
42.8 42.8 0
57.2 57.2 103
104
337
105
6822
0
0 150
200
100
100
50
51.4 51.4
48.6 103
104
323
105
5336
PBL
0
50.7 0
49.3 103
104
105
0
50.5 0
49.5 103
104
105
KLRG-1
Supplemental Figure 1 Similar KLRG-1 expression on antigen-specific CD8 T cells using ex vivo and in vivo staining procedures. The KLRG-1 expression on the antigen-specific effector cells was analyzed in P14 chimeric mice at day 4.75 after LCMV infection in the spleen, lymph nodes, PBMCs and the liver. The mice were killed, their tissues were isolated and the antigen-specific CD8 T cells were stained ex vivo for the gating markers CD8 and Thy1.1 at 2 hours after the in vivo injection of PBS or the KLRG-1 antibody. In the case of the ex vivo staining procedure, in which PBS was injected instead of the KLRG-1 antibody, the KLRG-1 antibody was included along with the CD8 and Thy1.1 antibodies. Representative histogram plots gated on CD81Thy1.11 P14 cells from two independent experiments are presented.
Cellular & Molecular Immunology: doi:10.1038/cmi.2014.48
Day 4 24.1
Day 8
Day 15
52.3
54.9
Day 60 24.6
Spleen
22.5
15
5.76
1.74
LN
33.9
67.4
69.3
22.9
40.1
22.1
51
65
Lung
28.4
16.9
Liver
39.4
20.5
PBL
KLRG-1
Supplemental Figure 2 Preferential localization of the KLRG-1int effector cells to the lymph nodes during CD8 T-cell responses to acute infections. P14 chimeric mice containing 13105 DbGP33-specific TCR-transgenic CD8 T cells were infected with LCMV. The mice were killed at the indicated days after infection, and the KLRG-1 expression on the gated antigen-specific CD8 T cells was analyzed directly ex vivo in both secondary lymphoid and non-lymphoid tissues. Representative FACS plots from three independent experiments are presented.
Cellular & Molecular Immunology: doi:10.1038/cmi.2014.48
Day 0
2hrs
RESEARCH ARTICLE IP
Isolate T cells from tissues
Stain for KLRG-1 ex vivo
Isolate T cells from tissues
KLRG-1 stained in vivo
IV
LCMVArm
PBS 2hrs
IP
IV
LCMVArm
KLRG-1 APC
in vivo 500
ex vivo 400
245
7643
400
233
4747
300
Cellular & Molecular Immunology advance online publication, ; doi:10.1038/cmi.2014.48 300 SPL
200
200
Supplementary Information accompanies100 the paper on Cellular & Molecular Immunology’s website 0(http://www.na49.6 50.4 0 10 10 10 ture.com/cmi). 5064 250 365 Supplementary Figure 1 3
4
Supplementary Figure 2
100
5
0
58.5 0
400
41.5 103
104
354
105
4317
200 300
iLN
150 200
100
100
50 0
0 1500
33.7 33.7
66.3 103
104
318
105
7534
0
0 1000
35.4 35.4
64.6 103
104
340
105
5709
800
1000
600
LVR
400
500
200
0
300
42.8 42.8 0
57.2 57.2 103
104
337
105
6822
0
0 150
200
100
100
50
51.4 51.4
48.6 103
104
323
105
5336
PBL
0
50.7 0
49.3 103
104
105
0
50.5 0
49.5 103
104
105
KLRG-1
Supplemental Figure 1 Similar KLRG-1 expression on antigen-specific CD8 T cells using ex vivo and in vivo staining procedures. The KLRG-1 expression on the antigen-specific effector cells was analyzed in P14 chimeric mice at day 4.75 after LCMV infection in the spleen, lymph nodes, PBMCs and the liver. The mice were killed, their tissues were isolated and the antigen-specific CD8 T cells were stained ex vivo for the gating markers CD8 and Thy1.1 at 2 hours after the in vivo injection of PBS or the KLRG-1 antibody. In the case of the ex vivo staining procedure, in which PBS was injected instead of the KLRG-1 antibody, the KLRG-1 antibody was included along with the CD8 and Thy1.1 antibodies. Representative histogram plots gated on CD81Thy1.11 P14 cells from two independent experiments are presented.
Cellular & Molecular Immunology: doi:10.1038/cmi.2014.48
Day 4 24.1
Day 8
Day 15
52.3
54.9
Day 60 24.6
Spleen
22.5
15
5.76
1.74
LN
33.9
67.4
69.3
22.9
40.1
22.1
51
65
Lung
28.4
16.9
Liver
39.4
20.5
PBL
KLRG-1
Supplemental Figure 2 Preferential localization of the KLRG-1int effector cells to the lymph nodes during CD8 T-cell responses to acute infections. P14 chimeric mice containing 13105 DbGP33-specific TCR-transgenic CD8 T cells were infected with LCMV. The mice were killed at the indicated days after infection, and the KLRG-1 expression on the gated antigen-specific CD8 T cells was analyzed directly ex vivo in both secondary lymphoid and non-lymphoid tissues. Representative FACS plots from three independent experiments are presented.
Cellular & Molecular Immunology: doi:10.1038/cmi.2014.48
