Bayan et al supplemental Figure 1

CD45+BrdU+DAPI

Supplemental Figure 1: Inflammatory cell infiltration inthe STZ treated mice. STZ-treated mouse pancreas werecostained with CD45 (leukocyte cell marker) (red), BrdU(green) and DAPI (blue). Top panel is the originalmagnification, 20X. Bottom panel is higher magnification ofthe corresponding IHC within the indicated square.

Bayan et al Supplemental Figure 2

d.

PtenloxP/loxP; RosalacZ/lacZ; RIP-Cre+

PtenloxP/loxP; RosalacZ/lacZ; RIP-Cre-

H&

EIn

Sit

u-

gal

sta

inin

g

Supplemental Figure 2: Generation of PtenloxP/loxP;Rosa26lacZ; RIPCre mice. Control (PtenloxP/loxP;Rosa26lacZ; RIPCre-) and Pten null (PtenloxP/loxP;Rosa26lacZ; RIPCre+) mice. pancreata were analyzed withH&E staining (top panel) and in situ β-gal staining (bottompanel). Blue stain indicates Pten-deleted β-cells. Originalmagnification, 20X.

Glu

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; B

rdU

; D

AP

IP

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Brd

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Bayan et al Supplemental Figure 3

a.

50 μm

Supplemental Figure 3: Proliferating activity are mostly confined to β-cells in non-treated mice. Pancreatic tissues were stained for glucagon, pancreatic polypeptide or somatostatin (Red) and costained with BrdU (Green) and DAPI (Blue) for nuclei. Very little BrdU positive cells are outside the islet confine (dotted circles). Right panels are a magnification of the area within the dashed squares.

Control Pten null

Bayan et al supplemental Figure 4

BrdU; Amylase; DAPI BrdU; Amylase; DAPI BrdU; Pan-CK; DAPIBrdU; Pan-CK; DAPI

A.

BrdU; Pdx-1 BrdU; GFAP; DAPIBrdU; Ngn3; DAPI BrdU; Nestin; DAPI

PtenloxP/loxP; Rosa26lacZ/lacZ; RIP-Cre+B.

50 μm

50 μm

Supplemental Figure 4: Exocrine or other stem/progenitor cell types do notcontribute to the extensive mitotic activity surrounding the islets in the STZtreated Pten null pancreas. (A) STZ-treated control (Pten+/+; Rosa26lacZ; RIPCre+)and mutant (PtenloxP/loxP; Rosa26lacZ; RIPCre+) pancreas were costained with either theacinar cell marker amylase or the ductal cell marker pan-cytokeratin (red) and BrdU(green) and DAPI (blue). The left two panels are the controls and the right two panelsare the mutants. Original magnification, 20X. (B) Mutant pancreas (PtenloxP/loxP;Rosa26lacZ; RIPCre+) were costained with various potential progenitor cell markers:pancreatic progenitor cell markers Ngn3 and PdX-1, neuronal stem cell marker nestinand astrocyte cell marker GFAP (red), and BrdU (green) and DAPI (blue). Top panel isthe original magnification 20X. Bottom panel is higher magnification of thecorresponding IHC. Dashed circles indicate islet area.

Control Pten null