A
B
C
0
5000
10,000
15,000
20,000 MT1: CT26
0
2000
4000
6000
8000
10,000 MT1: B16
0
2000
4000
6000 MT2: CT26
0
500
105
105
104
104
103
102 103
0
0100 102 103104 5
1000
1500
2000
2500 MT2: B16
TIL.DN TIL.SP TIL.DP
0
20,000
40,000
60,000
80,000
100,000
0
10
20
30
40
Zinpyr-1 MFI
Free Zn in CD8+ TILs
WT: TIL CD8 MT–/–: TIL CD8
10.6 29.2
WT MT–/–
GrzmB
EffMem
DP (Tim3+PD1+)
SP (Tim3–PD1+)
DN (Tim3–PD1–)
WT
MT–/–
RE
RE
Granzyme B
% Granzyme B
+ CD8
+ TILs
CD8
***NS
Figure S2. Metallothionein Expression, Zinc Dysregulation, and Effector Function in CD8+TILs, Related toFigure 2
(A) Expression of MT1 and MT2 as determined by qPCR in sorted CD8+TILs isolated from mice bearing CT26 colon carcinoma and B16 melanoma tumors.
(B) WT andMT/TILs were stained with Zinpyr-1 for measuring free Zn followed by cell surface staining and analyzed by flow cytometry. C) Granzyme
expression in WT andMT/CD8+TILs.