a
Luc-mRNAVEGF-C-mRNA(^103791113)
10104
1056
10107
8
Days
Cell numbe
rMeninges macrophage
(^103791113)
10104
1056
10107
8
Days
Cell numbe
rMeninges neutrophil
(^103791113)
10104
1056
10107
8
Days
Cell numbe
rMeninges Macrophage 2
(^103791113)
10104
1056
10107
8
Days
Cell number
Meninges pDC
(^103791113)
10104
1056
10107
8
Days
Cell number
Meninges cDC
(^103791113)
10104
1056
10107
8
Days
Cell number
Meninges ly6c-
(^103791113)
10104
1056
10107
8
Days
Cell number
Meninges ly6c+
(^103791113)
10104
(^105)
(^106)
7
Days
Cell numbe
r LN neutrophil
P=0.015
(^103791113)
101045
10106
1078
Days
Cell number
LN ly6c+
P=0.009
(^101791113)
102
103
Days
Cell number
LN resident DC
(^103791113)
104
105
Days
Cell number
LN migratory DC
(^104791113)
105
Days
Cell numbe
r LN pDCP=0.0003
(^104791113)
105
Days
Cell number
LN cDC
(^103791113)
104
105
106
Days
Cell numbe
r LN macrophage
(^104791113)
105
106
Days
Cell numbe
r
Brain neutrophil
P=0.04
(^104791113)
105
106
Days
Cell numbe
r
Brain pDC
P=0.003
(^103791113)
10104
1056
10107
8
Days
Cell numbe
r
Brain ly6c-
(^10579111315)
106
107
Days
Cell numbe
r Brain ly6c+P=0.037
(^10379111315)
101045
10106
1078
Days
Cell numbe
r Brain macrophage
(^10379111315)
104
105
106
107
Days
Cell numbe
r Brain cDC1
(^10379111315)
101045
10106
1078
Days
Cell numbe
r Brain microglia
Luc-mRNA VEGF-C-mRNA
FSC-A
SSC
FSC-H
FSC-W
SSC-WSSC-H
LiveCD45-BUV737
/dead-AmCy
an
CD19,CD3, NK1.1 BVCX3CR1-BV421 -605
CD11b-PELy6G-APC-Cy7
CD11b-PECD64-APC
Brain/Meninges
Lymph Node
M
Microglia/Meningeal M
Ly6G-APC-Cy7
Ly6C-A488
Other
Ly6C+ Monocytes
Ly6C- cells
Neutrophil
CX3CR1 low
CX3CR1 hi
MHCII-A700
CD11c-PE-Cy7
CD11c-PE-Cy7MHCII-A700
pDCResDC
MigDC
cDC
Other
Brain/Meninges
Lymph Node
CX3CR1 low
CX3CR1 hi
Other
All cells
Leukocytes
Single cell-1
Single cell-1
Single cell-2
Single cell-2
CD45+
CD45+
pDC
pDC
CD11cB220-BV496
b
c
d
e
f
g
Luc-mRNAVEGF-C-mRNA
Luc-mRNAVEGF-C-mRNA
(^50000)
1000015000
2000025000
MFI
Brain d11 MHCII
0
1000
2000
3000
MFI
Brain d11 CD80
ly6c+ monoly6c- mono
cDCpDC
macroph
agemicroglia
0
10000
20000
30000
40000
MFI
Brain d14 MHCII
ly6c+
monoly6c- monocDCpDC
macroph
agemicroglia
(^10000)
2000
30004000
5000
MFI
Brain d14 CD80
0
1000020000
3000040000
50000
MFI
LN d11 MHCII
0
1000
2000
3000
4000
MFI
LN d11 CD80
ly6c+
mono
ly6c-
monocDCpDC
macrophage
0
20000
40000
60000
MFI
LN d14 MHCII
ly6c+ mo
no
ly6c- mo
nocDCpDC
macrop
hage
0
1000
2000
3000
4000
MFI
LN d14 CD80
0
10000
20000
30000
40000
MFI
Meninges d11 MHCII
0
1000
2000
3000
MFI
Meninges d11 CD80
ly6c+ monoly6c- m ono
cDCpDC
macrophage
1
macrophage
(^0 2)
1000200000
3000400000
50000
MFI
Meninges d14 MHCII
ly6c+ mo
no
ly6c- mo
nocDCpDC
macroph
age 1
macrophage 2
0
1000
2000
3000
4000
MFI
Meninges d14 CD80
Extended Data Fig. 8 | Flow cytometry analysis of myeloid cell populations
after treatment with VEGF-C. Mice bearing 7-day tumours were treated with
Luc mRNA or VEGFC mRNA and evaluated for changes in the populations of
myeloid cells. a, Gating strategy for different myeloid cells. b–d, Cell counts of
different cell types were measured at different time points after VEGFC mR NA
treatment. cDC, classical dendritic cell; DC, dendritic cell; pDC, plasmacytoid
dendritic cell. e–g, MHC II and CD80 MFI levels were quantified and showed no
significant alteration after treatment with VEGFC mR NA. b and e are leukocytes
from brain tissue; c and f are leukocytes from draining cervical lymph nodes;
d and g are leukocytes from meninges (n = 3, 3 mice were pooled for each
replicate). Data are mean ± s.e.m P < 0.05; P < 0.01; P < 0.001; ****P < 0.0001
(two-tailed unpaired Student’s t-test).