Tanaka et al.
Supplementary Figure Legends
Supplementary Figure S1 Immunohistochemical analysis of EVI1 in pancreatic neoplastic tissues. A-J, Immunohistochemical analysis of EVI1 in pancreatic tissue ([A, C, E, G, I] H&E
staining; [B, D, F, H, J] EVI1 immunostaining). A-B, Acinal-ductal metaplasia. Ductal
metaplasia of acinar cells shows weak or absent expression of EVI1. C-D, PanINs. In the
PanIN-3 lesion (high-grade PanIN), EVI1 is expressed strongly in the nucleus. E-H, IPMNs.
In the IPMN lesion (both gastric-type and intestinal-type), EVI1 is expressed strongly in the nucleus. I-J, PDACs. In poorly differentiated PDACs, EVI1 is weakly or moderately
expressed.
Supplementary Figure S2 Oncogenic functions of EVI1 in pancreatic cancer cells. A-B, The effects of EVI1 knockdown on proliferation of EVI1-positive pancreatic-lineage
cells, HPDE (A) and KLM-1 cells (B). Growth assay was carried out in 96-well plates where
cells were plated at 3,000 cells per well and grown in 0.5% FBS medium. Experiments were
carried out in triplicate. * p < 0.01. C, The effects of EVI1 knockdown on cellular migration of PK-1 cells. * p < 0.01.
Supplementary Figure S3 Effects of EVI1 knockdown on other signaling pathways.
A, The effects of EVI1 knockdown on TGF-β signaling in HPDE cells. After transfection with control siRNA or siEVI1, HPDE cells were treated with or without TGF-β, and subjected to qRT-PCR analysis. B, The effects of EVI1 knockdown on p38 MAPK, Akt, and JNK pathway in PK-8, HPDE, and BxPC-3 cells. C, EVI1 knockdown decreased the expression level of Tanaka et al.
KRAS protein in BxPC-3 cells. D, The effects of EVI1 knockdown on KRAS mRNA expression in PK-8 cells. * p < 0.01. E, Phosphorylation of ERK was decreased by knockdown of EVI1 in BxPC-3 cells. F, EVI1 knockdown elevated p27Kip1 expression in
BxPC-3 cells. G, Expression changes of miRNA-96 in HPDE cells were analyzed using qRT-
PCR after EVI1 knockdown (72 h). * p < 0.01. H, miR-96 introduction suppressed the level of KRAS protein in BxPC-3 cells.