Anti γtubulin clone gtu 88

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)}.

Immunoblotting (IB) and signal detection was performed as previously described^{53 (link)}. The following antibodies were used for IB: anti-KIF21A (orb184767, Biorbyt, diluted 1:1000), anti‐γTubulin (clone GTU‐88, Sigma Aldrich, diluted 1:5000) and respective HRP‐conjugated antibodies (DAKO, diluted 1:5000).

The following antibodies were used for immunofluorescence: mouse anti α-tubulin (clone DM1A, Sigma-Aldrich) 1:1000; rabbit anti-Msps rb2219 (this study) 1:1000; rabbit anti-Cnn (Bettencourt-Dias et al., 2005 (link)) 1:2000; chicken anti-Dplp (Rodrigues-Martins et al., 2007 (link)) 1:1000; rabbit anti-mCherry (Abcam ab167453) 1:2000; mouse anti-LAMP1 human (clone H4A3, DSHB) 1:500; anti γ-tubulin (clone GTU-88, Sigma-Aldrich) 1:1000, rabbit anti-Rab5 antibody - Drosophila Early Endosome Marker (Abcam ab31261) 1:100; Recombinant rabbit monoclonal Anti-hRab5 antibody (Abcam ab218624) 1:1000
A rabbit anti-Msps was generated against the 1350-1785 amino acid fragment of Msps. DNA encoding the fragment was amplified by PCR from cDNA (Table S1), cloned in the pDONR221 entry vector, and recombined with the pDEST17 destination vector to generate a recombinant DNA encoding a 6xHis N-terminally tagged fusion fragment. The 6xHis fragment was expressed in bacteria, affinity purified on Ni-NTA Agarose beads (Qiagen) and used to for rabbit immunizations (Harlan UK). The final bleed was used for immunostainings and western blots.