Protein Expression Analysis in Cell Lines

Total cell lysates and cytoplasmic and nuclear protein fractions were extracted from cell cultures as described previously [51 (link)]. Standard Western blot analysis of protein expression was carried out using primary anti-HA (rabbit; Cell Signaling Technology), anti-TAZ (rabbit; Cell Signaling Technology), anti-YAP (rabbit; Cell Signaling Technology), anti-E-cadherin (mouse; BD Biosciences), anti-vimentin (rabbit; Cell Signaling Technology), anti-Merlin (rabbit; Santa Cruz Biotechnology), anti-LATS1 (rabbit; Cell Signaling Technology), anti-MST1 (rabbit; Cell Signaling Technology), anti-phosphorylated LATS1 (Thr1079, rabbit; Cell Signaling Technology), anti-phosphorylated MST1 (Thr183)/MST2 (Thr180, rabbit; Cell Signaling Technology), anti-TEAD (rabbit; Cell Signaling Technology), and anti-CTGF (mouse; Santa Cruz Biotechnology) antibodies. Equal protein-sample loading was monitored using an anti- glyceraldehyde-3-phosphate dehydrogenase (GAPDH) antibody for total cell protein lysates (rabbit; Santa Cruz Biotechnology), an anti-α-tubulin antibody for cytoplasmic fractions (mouse; Oncogene), and an anti-lamin B1 antibody for nuclear fractions (goat; Santa Cruz Biotechnology).

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Xie D., Cui J., Xia T., Jia Z., Wang L., Wei W., Zhu A., Gao Y., Xie K, & Quan M. (2015). Hippo transducer TAZ promotes epithelial mesenchymal transition and supports pancreatic cancer progression. Oncotarget, 6(34), 35949-35963.

Publication 2015

anti-HA anti-TAZ anti-YAP anti-E-cadherin anti-vimentin anti-Merlin anti-LATS1 anti-MST1 anti-TEAD anti-CTGF anti-lamin B1 antibody

Anti antibody Anti nuclear antibody Antibodies Cadherin Cell Cell cultures Ctgf Cytoplasmic Glyceraldehyde 3 phosphate dehydrogenase Goat Lamin b1 Lats1 Merlin Mouse Nuclear protein Oncogene Protein Rabbit Vimentin Western blot analysis α tubulin

Corresponding Organization : The University of Texas MD Anderson Cancer Center

Other organizations : Shanghai East Hospital, Second Military Medical University, Changhai Hospital

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Variable analysis

independent variables

Cell culture conditions

dependent variables

Protein expression levels of HA, TAZ, YAP, E-cadherin, vimentin, Merlin, LATS1, MST1, phosphorylated LATS1 (Thr1079), phosphorylated MST1 (Thr183)/MST2 (Thr180), TEAD, and CTGF

control variables

Equal protein-sample loading monitored using GAPDH for total cell protein lysates, α-tubulin for cytoplasmic fractions, and lamin B1 for nuclear fractions

controls

No positive or negative controls were explicitly mentioned

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