Nature 2020 01 30 Part.02

Article

Brain Tumor

0

50

100

150

Mean ER

V

a

row min row max

id SRR579545 (b6J)SRR579546 (b6J)SRR2927735 (b6J)SRR594402 (b6J)SRR3351236 (b6J)SRR2133109 (GL261)SRR2133110 (GL261)SRR2133111 (GL261)SRR2133112 (GL261)SRR3063568 (GL261)SRR3063569 (GL261)SRR3063566 (GL261)SRR3063567 (GL261)

Mpmv9Mpmv13

Mpmv2Pmv1

Mpmv1Xmv19

Mpmv11Pmv12

Mpmv12Mpmv5

Mpmv4Mpmv10

Pmv24Pmv22

Mpmv3Pmv17

Pmv8Pmv11

Pmv15Mpmv6

Pmv14Mpmv8

Pmv6

Pmv2Pmv23

Xmv12Pmv4

Xmv13Xmv42

Xmv8Pmv18

Pmv19Pmv21

Pmv20Emv2

Xmv41Xmv43

Pmv13Pmv9

Pmv5Pmv10

Pmv7

Mpmv7Xmv9

Xmv16Pmv16

Xmv10Xmv17

Xmv15Xmv18

id

row max

Xmv15Xmv1 8

Naive Mice

inguinal lymph node

0.48%

GL261 flank

inguinal lymph node

4.55%

PE-Empty

APC-Tetramer MHC I

d

Pmv20

Emv2

Xmv41

Xmv43

Healthy Brain GL261

b c beads2.99

Lymphocytes44.4 Single Cells89.1

50K 150K 250K

0

50K

100K

150K

200K

250K

CD3+60.9

0 104

0

-10^3

103

104

105

cd433.9

cd858.4

0 104

0

-10^3

103

104

105

CD44+63.5

0 104

0

-10^3

103

104

105

1.30tet

0 104

0

-10^3

103

104

105

5

0

50K

FSC-W

FSC-A

0

-10^3

CD3-BV605

CD45-APC-Cy 7

• 103
  CD4-PB
  CD8-BV7 11


• 103
  CD45-

APC-Cy7

CD44-PE

0

• 103
  CD45-APC-Cy7
  APC-Tetramer MHC I

Extended Data Fig. 6 | Validation of endogenous retrovirus EMV2 as a
tumour antigen for GL261. a, RNA-seq analysis of mouse endogenous
retrovirus elements in publicly available datasets on C57BL/6J mice brains and
GL261 cell lines from various sources. b, Quantification of endogenous
retrovirus elements in healthy brain and tumour tissue, from RNA-seq
(brain, n = 9; tumour, n = 6). c, Gating strategies for tetramer staining.

d, Mice were injected with 500,000 GL261 cells or PBS in the f lank. Seven days

after tumour inoculation, draining inguinal lymph nodes were collected and

emv2-env (Kb-restricted peptides aa 604–611 of p15E protein (KSPWFTTL))

tetramers were used to validate tumour-specific T cell proliferation. The

experiments were repeated twice independently with similar results. Data are

mean ± s.e.m.