Western Blot Analysis of Hippo-YAP Pathway

Jejunal tissues and cultured cells were lysed, and the extracted proteins were analyzed. The primary antibodies used for Western blot were rabbit anti-Mst1 (1:1000; Cell Signaling), anti-p-Mst (1:1000; Cell Signaling), anti-Lats1 (1:1000; Cell Signaling), anti-p-Lats (Yu et al. 2010 (link)), anti-YAP (1:1000 [Cell Signaling, #4912], 1:1000 [Santa Cruz Biotechnology, H-9 and 63.7], and 1:1000 [Novus, NB110-58358]), anti-p-YAP (S127; 1:1000; Cell Signaling), anti-p-YAP (S381; 1:1000; Cell Signaling) (Kim et al. 2013 (link)), anti-β-catenin (1:2500; Sigma), anti-YAP/TAZ (1:1000; Cell Signaling), anti-Axin2 (1:1000; Cell Signaling), mouse anti-APC (1:200; Calbiochem, Ab-1), anti-Flag (1:5000; Sigma, M2), anti-HA (1:5000; Sigma), anti-Myc (1:2000; Calbiochem), anti-Actin (1:5000; Millipore), and rat anti-Flag (1:2000; BioLegend, L5). Signals were quantified by ImageJ.

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Cai J., Maitra A., Anders R.A., Taketo M.M, & Pan D. (2015). β-Catenin destruction complex-independent regulation of Hippo–YAP signaling by APC in intestinal tumorigenesis. Genes & Development, 29(14), 1493-1506.

Publication 2015

rabbit anti-Mst1 anti-p-Mst anti-Lats1 anti-p-Lats anti-YAP NB110-58358 anti-p-YAP anti-β-catenin anti-YAP/TAZ anti-Axin2 anti-Flag anti-HA anti-Actin rat anti-Flag

Actin Antibodies Axin2 Cultured cells Jejunal Lats Lats1 Mouse Novus Proteins Rabbit Tissues Western blot β catenin

Corresponding Organization :

Other organizations : Johns Hopkins University, Johns Hopkins Medicine, Howard Hughes Medical Institute, The University of Texas MD Anderson Cancer Center, Kyoto University

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Variable analysis

independent variables

Not explicitly mentioned

dependent variables

Protein expression and phosphorylation levels of Mst1, Lats1, YAP, and related signaling proteins measured by Western blot

control variables

Not explicitly mentioned

controls

Positive control: None mentioned
Negative control: None mentioned

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