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2 protocols using phospho lats1 s909

1

Comprehensive Protein Analysis Protocol

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Cells were lysed in SDS lysis buffer (240 mM Tris-acetate, 1% SDS, 1% glycerol, 5 mM EDTA pH 8.0) with DTT, protease inhibitors, and a cocktail of phosphatase inhibitors. Antibody against vimentin, and MCP-1 were purchased from Abcam (Cambridge, MA, USA). Antibodies detecting CD44 and phospho-YAP S127 were purchased from GeneTex, Inc. (Irvine, CA, USA). Antibodies against YAP, phospho-MST1/MST2 T183/T180, MST1, phospho-LATS1 S909, LATS1, CYR61, CTGF, phospho–AKT T308, AKT, and phospho–GSK3β S9, phospho-ERK1/2, ERK1/2 were purchased from Cell Signaling Technology (Danvers, MA, USA). The β-actin, GAPDH, and α -tubulin antibodies were purchased from Novus Biologicals (Littleton, CO, USA). Antibody against E-cadherin was purchased from BD (Franklin Lakes, NJ, USA).
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2

Reagents for Protein Interaction Studies

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Tamoxifen (catalog #T5648), 4-hydroxy Tamoxifen (catalog #H7904), MG132 (catalog #M8699), N-ethylmaleimide (NEM) (catalog #E3876), cycloheximide (catalog #239763), reduced L-glutathione (catalog #G4251) and D-maltose (catalog #M9171) were purchased from Sigma-Aldrich. Antibodies directed against ERG (catalog #ab92513), CD31 (catalog #ab7388), FAT1 (catalog #ab109202), FAT4 (catalog #ab130076) were from Abcam. Isolectin GS-IB4 (catalog #I21413/I32450) was from Life Technologies GmbH. Antibodies directed against HA (catalog #H6533), Flag M2 (catalog #A8592) were from Sigma-Aldrich. Antibodies directed against YAP/TAZ (catalog #8418), YAP (catalog #4912), Lamin A/C (catalog #2032), GAPDH (catalog #2118), LATS1 (catalog #3477), phospho-LATS1 (S909) (catalog #9157), phospho-LATS1 (catalog #1079) (catalog #8654), phospho-MST1 (T183) (catalog #3681) phospho-MOB1 (T35) (catalog #8699) Ubiquitin (catalog #3936S), Cleaved Caspase-3 (catalog #9664), CYLD (catalog #8462) were obtained from Cell Signaling. The MIB2 antibody (catalog #A301-414A) was from Bethyl Laboratories, and Matrigel (catalog #356231) was purchased from Merck. Isopropyl-β-D-1-thiogalactopyranosid (IPTG) (catalog #CN08.2) and imidazole (catalog #X998.3) were from Carl Roth GmbH.
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