SUPPLEMENTARY DATA
Supplementary Figure 1. Bim and Puma upregulation in Pdx1+/- mouse islets. A-B: Bim and Puma
expression levels assessed by immunofluorescence in islets from wild type and Pdx1+/- mcie fed HFD
for 15 weeks.
A
Bim/Insulin/Nucleus
Pdx1 +/+
Pdx1 +/-
B
Puma/Insulin/Nucleus
Pdx1 +/+
Pdx1 +/-
©2013 American Diabetes Association. Published online at http://diabetes.diabetesjournals.org/lookup/suppl/doi:10.2337/db13-1513/-/DC1
SUPPLEMENTARY DATA
Supplementary Figure 2. Bim and Puma KD inhibit β-cell apoptosis induced by Pdx1 suppression. AC: Pdx1 KD increased cleaved caspase3 protein levels (A). Bim or Puma KD inhibits an increase of
cleaved casaspe3 protein levels induced by Pdx1 KD in MIN6 cells (B-C). 3 days after Pdx1 KD or
Bim/Pdx1or Puma/Pdx1 DKD in MIN6 cells, immunoblot of Pdx1, Bim, Puma and cleaved caspase3
in MIN6 cells. D: Measurement of Cell death. 4 days after Bim/Pdx1 DKD or Puma/Pdx1 DKD in
MIN6 cells, cell death was determined by PI-staining (n=3). Values are mean ± SEM. ***P < 0.001 vs.
control cells. # P < 0.05 vs. Pdx1KD cells.
A
B
Pdx1 KD
-
+
-
+
+
+
Pdx1 KD
Bim KD
+
-
Cleaved casp3
Pdx1
BimEL
Cleaved casp3
BimL
Actin
Bims
Pdx1
Actin
-
+
-
+
+
+
Pdx1 KD
Puma KD
Cleaved casp3
Puma
Pdx1
Actin
D
Cell death (PI staining)
C
80
60
***
#
40
#
20
0
©2013 American Diabetes Association. Published online at http://diabetes.diabetesjournals.org/lookup/suppl/doi:10.2337/db13-1513/-/DC1
SUPPLEMENTARY DATA
Supplementary Figure 3. The characterization of the high fat diet on the mice. A-B: Body weight of
the mice feeding high fat diet. Values are thes mean ± SEM, n=6-14. C-D: Feeding glucose levels of
the mice at age of 16 weeks. Values are thes mean ± SEM, n=8-12. *P < 0.05 and ***P < 0.001 vs.
wild type mice. ### P < 0.001 vs. Pdx1+/- mice. E-F: Fasting glucose levels of the mice at age of 16
weeks. Values are thes mean ± SEM, n=10-16. *P < 0.05 and ***P < 0.001 vs. wild type mice. ### P
< 0.001 vs. Pdx1+/- mice.
A
B
40
35
35
30
30
Body weight (g)
Body weight (g)
40
25
20
Pdx1 +/+Puma +/+
15
Pdx1 +/-Puma +/+
Pdx1 +/+Puma -/Pdx1 +/-Puma -/-
10
5
25
20
10
5
0
0
0
2
4
6
8
Weeks (HFD)
10
0
12
D
***
350
300
###
250
200
*
150
100
50
12
***
###
250
*
200
150
100
0
Pdx1 +/Puma -/F
200
Pdx1 +/+
Bim+/+
200
150
###
*
100
125
0
Pdx1 +/+ Pdx1 +/- Pdx1 +/+
Puma +/+ Puma +/+ Puma -/-
Pdx1 +/Puma -/-
Pdx1 +/- Pdx1 +/+ Pdx1 +/Bim+/+ Bim-/- Bim-/-
***
***
Blood Glucose(mg/dl)
Blood Glucose(mg/dl)
10
50
Pdx1 +/+ Pdx1 +/- Pdx1 +/+
Puma +/+ Puma +/+ Puma -/-
50
4
6
8
Weeks (HFD)
300
0
E
2
350
Blood glucose(mg/dl)
Glood Glucose(mg/dl)
C
Pdx1 +/+Bim+/+
Pdx1 +/-Bim+/+
Pdx1 +/+Bim-/Pdx1 +/-Bim-/-
15
150
###
100
50
0
Pdx1 +/+
Bim+/+
Pdx1 +/Bim+/+
Pdx1 +/+ Pdx1 +/Bim-/- Bim-/-
©2013 American Diabetes Association. Published online at http://diabetes.diabetesjournals.org/lookup/suppl/doi:10.2337/db13-1513/-/DC1