Embryos were fixed and stained as described by Dent et al.31 . The mouse monoclonal anti-acetylated α-tubulin antibody was used to visualized ciliated cells32 (link). The rabbit anti-Xenopus Cetn antibody (anti-XlCetn) was supplied by Sergie Sokol20 (link). The rabbit anti-human Cetn1 antibody (anti-HsCetn1) was purchased from Sigma. The anti-HsCetn1 antibody is directed against the C-terminal 15 amino acids of Cetn1; a similar sequence exists at the C-termini of X. laevis Cetn2 (13/15 identical) and Cetn4 proteins (12/15 identical), but is absent from the Cetn3 (5/15 identical)(data not shown). Notochord was visualized using the mouse monoclonal anti-keratin sulfate antibody MZ15 obtained from the Developmental Studies Hybridoma Bank33 (link). Immunoperoxidase-stained explants were bleached before staining while immunofluorescently-labeled embryos were not. Fluorescent images were collected using a Zeiss 510 Confocal Laser Scanning Microscope.
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