Download Supplementary Figure 1-12

Supplemental Figures
Figure S1. Validation of miRNA microarray data by qRT-PCR in the test cohort (n=69)
(Related to Figure 1).
Test Cohort (NanoString, n=69)
A
C
B
Kaplan-Meier analysis
miR-301b
miR-15b
miR-10b
miR-93
miR-590-5p
Correlation with qRT-PCR value
Figure S2. MIF regulates miR-301b expression in human pancreatic cancer
A
BxPC-3
MiaPaCa-2
Panc 10.05
SU.86.86
CFPAC-1
Panc-1
Capan-1
Capan-2
C
B
MiaPaCa-2
D
Panc-1
MiaPaCa-2
Capan-2
Capan-2
E
Panc-1
MIF
β-Actin
Capan-2
Capan-2
Figure S3. NR3C2 is a potential target of miR-301b (Related to Figure 2).
A
Gene Ontology analysis of candidate targets of miR-301b
qRT-PCR
Panc-1
B
Capan-2
C
pmiR-Luc-NR3C2 WT
Firefly pmiR
1367-1388
Firefly pmiR
1736-1758
pmiR-Luc-NR3C2 MUT
Firefly pmiR
×
1736-1758 MUT
Sequencing of pmiR-Luc-NR3C2 WT and Mutant reporter clones
D
E
293T
Figure S4. Endogenous NR3C2 expression was negatively correlated with MIF and miR-301b in a panel
of human pancreatic cancer cell lines (Related to Figure 2).
A
a (MiaPaCa-2)
BB
b (SU.86.86)
h
g
f
d
d (CFPAC-1)
e
h
c (Panc-1)
e (Capan-1)
g
e
f
d
f (Panc 10.05)
c
b
a
c
g (Capan-2)
h (BxPC-3)
a
b
Figure S5. NR3C2 enhances sensitivity to gemcitabine in pancreatic cancer cells (Related to
Figure 3).
A
NR3C2
NR3C2
β-Actin
β-Actin
B
C
Panc-1
Control
MiaPaCa-2
NR3C2
Control
NR3C2
Figure S6. Knockdown of NR3C2 increases proliferation, migration and invasion, and
decreases the sensitivity of pancreatic cancer cells to gemcitabine (Related to Figure 3).
B
A
NR3C2
β-Actin
Capan-2
sh-Control
sh-NR3C2
C
Migration
Invasion
Capan-2
D
Figure S7. A lower endogenous NR3C2 expression was associated with EMT phenotype
(Related to Figure 3).
A
NR3C2
E-cadherin
N-cadherin
β-Actin
B
Mophology
Panc-1
Capan-2
E-cadherin
E-cadherin/DAPI
Figure S8. NR3C2 inhibits epithelial-to-mesenchymal transition (EMT) (Related to Figure 3).
B
A
E-Cadherin
Merge
DAPI
sh-Control
Panc-1
sh-NR3C2
sh-Control
Capan-2
sh-NR3C2
C
D
Sh-NR3C2
Sh-Control
Panc-10.05
E-Cadherin
DAPI
Merge
E
sh-Control
Panc-10.05
sh-NR3C2
Figure S9. MIF-induced miR-301b expression was blocked by anti-miR-301b (Related to Figure 4).
Figure S10. MEK inhibitor AZD6244 didn’t alter miR-301b or NR3C2 in MIF-overexpressing
Panc-1 and Capan-2 cells (Related to Figure 5).
Figure S11. NR3C2 expression is decreased in tumors as compared to adjacent nontumor tissues
in PDAC (Related to Figure 7).
qRT-PCR
(Validation Cohort-2)
Affymetrix Gene Array
A
(Test Cohort)
B
Figure S12. A lower expression of NR3C2 is associated with poor survival in multiple
independent cohorts (Related to Figure 7).
TCGA Cohort
B
A
C
Moffitt Cohort
D