Molecular Mechanisms of Pancreatic Cancer Progression

Whole Pancreatic cancer cells were lysed on ice in a lysis buffer (RIPA, Beyotime, Shanghai, China) with a protease inhibitor mixture cocktail (Roche, Switzerland) after culturing in MRC-5-CM for 14 days. Western-blot analysis was performed by established protocols 43 (link). Anti-CTNNA1 and CTNNB1, anti-E-cadherin, anti-N-cadherin, anti-ZEB-1, ZEB-2, Snail, Slug and anti-vimentin primary antibodies were purchased from (Abcam); anti-MMP-2, 3, 13 and 21, anti-WNT-2, anti-WNT16, anti-TGFB1, anti-Src, P-Src-Y418 and P-Src-Y529 primary antibodies were purchased from (Epitomics); anti-Bcl-2, Bcl-xl, Bad, Bax, Bim, Bid, Caspase-3 and anti-β-actin were from ( Cell Signaling Technology).

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Ding S.M., Lu A.L., Zhang W., Zhou L., Xie H.Y., Zheng S.S, & Li Q.Y. (2018). The role of cancer-associated fibroblast MRC-5 in pancreatic cancer. Journal of Cancer, 9(3), 614-628.

Publication 2018

RIPA protease inhibitor mixture cocktail anti-E-cadherin anti-N-cadherin anti-ZEB-1 Snail anti-vimentin anti-WNT-2 anti-WNT16 anti-TGFB1 anti-Src P-Src-Y418 P-Src-Y529 anti-Bcl-2 Bcl-xl Caspase-3 anti-β-actin

Actin Anti antibodies Antibodies Bcl 2 Buffer Cadherin Caspase 3 Cells Ctnna1 Ctnnb1 Mmp 2 Pancreatic cancer Protease inhibitor Ripa Slug Snail Tgfb1 Vimentin Western blot analysis

Corresponding Organization : Blood Center of Zhejiang Province

Other organizations : Zhejiang University

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Variable analysis

independent variables

Culture in MRC-5-CM for 14 days

dependent variables

CTNNA1 and CTNNB1 expression
E-cadherin, N-cadherin, ZEB-1, ZEB-2, Snail, Slug, and vimentin expression
MMP-2, 3, 13, and 21 expression
WNT-2 and WNT16 expression
TGFB1 expression
Src, P-Src-Y418, and P-Src-Y529 expression
Bcl-2, Bcl-xl, Bad, Bax, Bim, Bid, and Caspase-3 expression

control variables

Amount of protein loaded for Western blot analysis (equal loading)

positive controls

Anti-β-actin antibody to verify equal loading

negative controls

Not explicitly mentioned

Annotations

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