Anti perm 3141

Immunofluorescence was performed as previously described (Dietrich and Hiiragi, 2007 (link)). The following antibodies and dilutions were used: monoclonal mouse anti-CDX2 (MU392-UC, BioGenex) 1:200, rabbit monoclonal anti-CDX2 (ab76541, Abcam) 1:200, mouse monoclonal anti-YAP (sc-101199, Santa Cruz Biotechnology) 1:200, rabbit polyclonal anti-pERM (3141, Cell Signalling) 1:250, rat monoclonal anti-E-Cadherin (U3254, Sigma) 1:250, mouse monoclonal anti-TEAD4 (ab58310, Abcam) 1:100, rabbit polyclonal anti-DsRed (632496 living colors Clontech) 1:400, goat polyclonal anti-GFP (R1091P, Acris, Origene) 1:200, rat monoclonal anti-HA (11867423001, Sigma) 1:200. Secondary Alexa Fluor conjugated antibodies (Life Technologies) were used at 1:1000. Nuclei were visualized by incubating embryos in DAPI at 1 μg/ml.

Zebrafish whole-mount immunofluorescence (IF) was performed as previously described^{33 (link)}. The following antibodies were used for IF: anti-CLIC5 (B-23; sc-133468; Santa Cruz Biotechnology; 1:10), anti-CLIC5 (A-11; sc-271863; Santa Cruz Biotechnology; 1:10), anti-γTubulin (clone GTU‐88; Sigma Aldrich; 1:5000), anti-GFP (ab13970; Abcam; 1:200), anti-acetylated αTubulin (clone 6-11B-1; Sigma Aldrich; 1:500) and anti-phospho-Ezrin (Thr567) (PA5-37763; Invitrogen; 1:200). Cy3 (1:1000) and Alexa-488/-546/-647 (1:1000) labelled secondary antibodies were purchased from Jackson Immunoresearch and Molecular Probes (Invitrogen), respectively. Immunoblotting (IB) was performed as previously described^{47 (link)}. The following antibodies were used for IB: anti-CLIC5 (B-23; sc-133468; Santa Cruz Biotechnology; 1:200), anti‐pERM (#3141; Cell Signaling; 1:1000), anti‐ERM (#3142; Cell Signaling; 1:1000), anti‐γTubulin (clone GTU‐88; Sigma Aldrich; 1:5000) and respective HRP‐conjugated antibodies (DAKO; 1:5000). Histological and transverse sectioning procedures were performed as previously described^{33 (link)}.