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Actin PEA-15 2774 Hey KOC-7c OVCA420 OVCA432 OVTOKO RMG-1 SKOV3.ip1 Supplementary Fig. S1. Expression level of endogenous PEA-15 in ovarian cancer cell lines by Western blotting analysis. A total of 15 g of proteins was used for SDS-PAGE. The expression levels of PEA-15 protein for each cell line were evaluated using LI-COR software. Supplementary Figure S1. Lee et al.
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B MMP-7 Vector PEA-15-AA PEA-15-DD A Supplementary Fig. S2. PEA-15-AA-expressing cells showed downregulation of -catenin target genes. To test the effect of PEA- 15-AA-induced- -catenin inhibition on -catenin target genes, we performed quantitative RT-PCR and immunohistochemical staining. (A) Quantitative RT-PCR analysis of c-myc and c-met. (B) Immunohistochemical staining of MMP-7 using xenograft tumor samples. Scale bars: 50 m. Supplementary Figure S2. Lee et al.
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Tumor tissue Cancer-adjacent normal tissue pPEA-15-S104 pPEA-15-S116 -catenin Supplementary Fig. S3. PEA-15 phosphorylated at Ser104 (pPEA-15-S104) and PEA-15 phosphorylated at Ser116 (pPEA-15- S116) were highly overexpressed in breast tumor tissues. To investigate the expression status of pPEA-15-S104, pPEA-15-S116, and b- catenin, we performed immunohistochemical staining using a human breast tumor microarray (US Biomax Inc). Original magnification, ×200 (insets, ×40). Supplementary Figure S3. Lee et al.
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Supplementary Figure S4. Lee et al. Supplementary Fig. S4. Effect of PEA-15s in an ovarian cancer xenograft model. (A). Upper table showed incidence of tumor formation of vector, PEA-15-WT, PEA-15-WT, PEA-15-DD-3, and PEA-15-AA-1 clone. Lower panel, tumor weights were significantly smaller with PEA-15-AA-1 clone than with vector (P<0.0001), PEA-15-WT (P=0.0039), and PEA-15-DD-3 (P<0.0001) clone. (B). Representative images showed tumor formation of SKOV3.ip1-vector, SKOV3.ip1-PEA-15-WT, SKOV3.ip1-PEA-15-DD-3 and SKOV3.ip1-PEA-15-AA-1 cells in mouse peritoneal area. White arrows indicate tumors. A B Total number of mice Incidence of tumor formation (%) Vector 1010/10 (100%) PEA-15-WT 1010/10 (100%) PEA-15-DD-3 1010/10 (100%) PEA-15-AA-1 88/10 (80%)
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Supplementary Table S1. Incidence of high and low -catenin expression levels in PEA-15-AA, PEA-15-DA, and PEA-15-DD tumor tissues. PEA-15 phosphorylation status -catenin expression PEA-15-AAPEA-15-DAPEA-15-DD High (intensity=2 or 3)2 (25%)7 (41.2%)11 (33.3%) Low (intensity=0 or 1)6 (75%)10 (58.8%)22 (66.7%) Total sample number81733 Supplementary Table S1. Lee et al.
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