TET3

Supplementary  informations                 A  miR-­‐150  /  TET3  pathway  regulates  the  generation  of  mouse  and  human  non-­‐ classical  monocyte  subset     Selimoglu-­‐Buet  D.  et  al.    

Selimoglu-Buet D et al, Supplementary Figure 1 (related to Figure 1) B

25

hsa-miR-150

20

hsa−miR−150

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8.5

11

8.0

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LMMC

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MUT_FLT3 CTRL MUT_RAS LMMC

MUT_CBL MUT_AML MUT_TET2 MUT_FLT3MUT_ASXL1 MUT_RAS MUT_CYTOK MUT_CBL MUT_TET CTRL LMMC MUT_AML MU

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CMML

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0.0001

0.0001

0.00001

0.00001

CTL CMML

CTL CMML

****

Data 1

10000

mir451/hprt rev

100

1000

10

100

1

10

0.1

1

0.01

0.1

0.001

0.01

0.0001

CTL CMML

CTL CMML

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C

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TL

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IR

iR

15

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CTL CMML

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*

100

0.0001

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M IR

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M IR

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1

1

TL

0.01 0.001

10

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100

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mir451 /cyclo rev

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15 0

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****

Data 1

M IR

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1

hsa-miR-451

M IR 15 0

1

hsa-miR-150

TL

** 100

D

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***

NS

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hsa-miR-494

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miR expression miR expression (relative to HPRT) (relative to PPIA)

hsa-miR-150 miR expression miR expression (relative to PPIA) (relative to RPL32)

● ●

●

MUT_AML

CTL CMML CTL

10

7.5

log2(I)

7.0 6.5

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5.5

5.0

CTRL

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log2(I)

7.5 7.0

log2(I)

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5.0

5.0

0

11.0 10.0 9.0 8.0

●

●

6.0

6.0 5.5

5

7.0

7.0 6.5

10

hsa−miR−451

8.0

6.0

Log2(I)

15

C

hsa-miR-494

hsa-miR-451

●

6.0

A

WBC 9 (10 /L) (10 WBC /L)

60 40

1

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1 1000.01

100

0.1

1

10

100

10

100 10 1 0.1 0.01 CMML-0 CMML-0

9

10

1

10

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G 0.1

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10

100

100

10

0.1

1

10

100

0.01

0.1

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hsa-miR-150 expression (/RUN6B)

10

100

CMML-1

CMML-1

CMML-2

CMML-2

1000 100

TET2

SRSF2

ASXL1

RAS

RUNX1

10 1 0.1 0.01

WT MT WT MT WT MT WT MT WT MT

2 Te wt t2 SR m SF ut SR 2 W SF T AS 2 m XL ut AS -1 XL WT -1 m RA ut S RA WT S AM m L- ut AM 1 W L- T 1 m ut

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0.1 10000.01

Platelets (10 /L)

0.1 200.01

1

Te t

1

9 9/L) Platelets (10

Hemoglobin (g/dL) Hb (g/dL)

10

Neutrophils 9/L) (10 PN (10 /L)

0.1

9

Monocytes 9/L) (10 Monocytes count (10 /L)

20 0 1000.01

100

9

Age (year) age (year)

80

lien LMMC1 ou2

miR-150hsa-miR-150 expression (relative to RNU6B) expression (/RUN6B)

F

1000

100

hsa-miR-150 expression (/RUN6B)

E

Supplementary  Figure  1.  Differen4al  miRNA  expression  profiles  in  healthy  controls  or  CMML   sorted  peripheral  blood  monocytes  and  rela4onship  with  clinical  and  biological  features.     A.  Hierarchical  clustering  of  miRNA  profiles  obtained  by  microarray  analysis  (learning  cohort),   established   using   the   Euclidean   distance   method   and   the   Ward   clustering   method,   and   validated  by  boostraping  (package  pvclust).  B.  Box  plot  showing  the  variaEon  in  hsa-­‐miR-­‐150,   hsa-­‐miR-­‐451  and  hsa-­‐miR-­‐494  expression  in  samples  of  the  learning  cohort  analyzed  in  figure   1A  (centre  line  :  median,  whiskers:  min  to  max).  C.  Box  plot  showing  the  relaEve  expression  of   hsa-­‐miR-­‐150,   hsa-­‐miR-­‐451   and   hsa-­‐miR-­‐494   as   measured   by   qRT-­‐PCR   in   CD14+   cell   samples   used  for  microarray  analysis  (learning  cohort).  Normalized  to  L32  or  PPIA  (CTL=5;  CMML=33),   centre   line   :   median,   whiskers:   min   to   max.   D.   qRT-­‐PCR   analysis   of   hsa-­‐miR-­‐150   and   hsa-­‐ miR-­‐451   expression   in   healthy   donor   and   CMML   paEent   CD14+   cells   of   controls   and   CMML   paEents   (validaEon   cohort).   hsa-­‐miR-­‐150   normalized   to   PPIA   (CTL=19;   CMML=124)   or   HPRT   (CTL=19;  CMML=143).  hsa-­‐miR-­‐451  normalized  to  PPIA  (CTL=24;  CMML=44)  or  HPRT  (CTL=24;   CMML=52).   Red   lines:   median   with   interquarEle   range.   Mann-­‐Whitney   test:   *   P

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