Immunofluorescence and Immunoblotting of Zebrafish

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)}.