Nature 2020 01 30 Part.02

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(^0) -10 4 0 104 105106
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(^0) -10 4 0 104 105106
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(^0) -10 4 0 104 105106
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AKT-phos-BV421
AKT-phos-BV421
Lymph node
Meninges
AAV-VEGF-C tumor + Luc-mRNA
LEC
BEC
LEC
BEC
LEC
BEC
LEC
BEC
LEC
BEC
LEC
BEC
tumor + VEGF-C-mRNA
AAV-VEGF-C tumor + Luc-mRNA tumor + VEGF-C-mRNA
AAV-VEGF-C tumor + Luc-mRNA tumor + VEGF-C-mRNA
AAV-VEGF-C tumor + Luc-mRNA tumor + VEGF-C-mRNA
FSC-A
SSC
FSC-W
FSC-H
SSC-W
SSC-H
CD45-BV605
FSC-H
CD31-A647
PDPN-PE
CD31-A647
PDPN-PE
Cells Cells
Single cell 1
Single cell 1
Single cell 2
Single cell 2
CD45-
CD45+
CD45- LEC CD45+
BEC
46.4% 12.6% 32.5%
2.63% 1.40% 2.86%
4.52% 0.82% 4.04%
1.65% 2.34% 2.04%
Baseline: MFI 1064, 9.82%
Baseline: MFI 313, 0.96%
Baseline: MFI 563, 2.27%
Baseline: MFI 349, 4.59%
MFI 2797 MFI 1072 MFI 1935
MFI 468 MFI 489 MFI 472
MFI 1064 MFI 563 MFI 663
0 MFI 337 MFI 333 MFI 341
1000
2000
3000
4000
5000
MFI
Meninges BEC
0
1000
2000
3000
4000
5000
MFI
Meninges LEC
1.6x
2.1x
P= 0.02
ns P= 0.058

• 0
  20
  40
  60
  80
  % AKT(pS473)+
  (CD45-CD31+PDPN+)
  Meninges LEC
  P= 0.01


• P= 0.03


• 
  

  2.4x
  3.2x
  P= 0.0007

  
  


• P= 0.02

  ---

  
  

  P= 0.004
  2.1x0.4x1.7x
  P= 0.06
  P= 0.007
  ns
  1.2x 1.2x
  WT
  AAV-VEGF-
  C
  tumor + Luc-mRNAtumor + VEGF-C-mRNA
  0
  20
  40
  60
  80
  % AKT(pS473)+ (CD45-CD31+PDPN-
  ) LN BEC
  0
  20
  40
  60
  80
  % AKT(pS473)+ (CD45-CD31+PDPN-
  ) Meninges BEC
  WT
  AAV-VEGF-C
  tumor + Luc-mRNA
  tumor + VEGF-C-mRNA
  0
  1000
  2000
  3000
  4000
  5000
  MFI
  LN BEC
  WT
  AAV- VEGF-C
  tumor + Luc-mRNAtumor + VEGF-C-mRNA
  0
  20
  40
  60
  80
  % AKT(pS473)+
  (CD45-CD31+PDPN+)
  LN LEC
  WT
  AAV-VEGF-C
  tumor + Luc-mRNAtumor + VEGF-C-mRNA
  0
  1000
  2000
  3000
  4000
  5000
  MFI
  LN LEC
  LYVE1CD31DAPI
  WT
  tumorLuc-mRNA
  tumor
  VEGF-C-mRNA
  tumor
  Luc-mRNA
  tumor
  VEGF-C-mRNA
  LYVE1CD31DAPI LYVE1CD31DAPI
  f
  g
  0102030405060
  0
  20
  40
  60
  80
  100
  Days elapsed
  Percent surviva
  l No tx
  -d60 AAV
  d0 AAV
  d7 AAV
  d0 mRNA
  d3 mRNA
  d7 mRNA
  P = 0.003
  P = 0.229
  P = 0.169
  P = 0.044
  P = 0.035
  P = 0.003
  **

  
  


• 
  
  
  
  • 
    
  
  
  
  


• 
  

  No tx
  d7 d0 AAAAVV
  d3 mRNAd0 mRNA
  d7 mRNA
  -d60 AAV
  d7 d14 d21d 28
  no tx
  d7 AAV
  d0 AAV
  d7 mRNA
  -d60 AAV
  d3 mRNA
  d0 mRNA
  h
  (^0051015202530)
  2104
  4104
  1106
  Days
  RLU
  (^0051015202530)
  2104
  4104
  1106
  Days
  RLU
  (^0051015202530)
  2104
  4104
  1106
  Days
  RLU
  (^0051015202530)
  2104
  4104
  1106
  Days
  RLU
  (^0051015202530)
  2104
  4104
  1106
  Days
  RLU
  N = 4
  (^0051015202530)
  2104
  4104
  1106
  Days
  RLU
  (^0051015202530)
  2104
  4104
  1106
  Days
  RLU
  (^0051015202530)
  2104
  4104
  1106
  Days
  RLU
  N = 1
  No tx
  -d60 AAV d0 AAV
  d7 AAV d0 mRNA
  d3 mRNA d7 mRNA
  GL261-Luc
  a b
  c
  de
  Extended Data Fig. 4 | See next page for caption.