ZFIN Image: Marsay et al., 2021, Fig 5

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Figure Caption

Fig 5

Generation of cd9a mutant in the cd9b ^pg15 KO background to create a double cd9 KO mutant.

A: cd9a mutant allele sequence from exon 3, showing CRISPR target sequence in blue. The 4bp deletion (dashes) and 12bp insertion (lowercase) results in an 8bp insertion as indicated under the WT sequence. This leads to a frameshift changing codon 61 from CTC (Leu) to GCG (Ala), then 9 aberrant amino acids (red lettering) followed by a stop codon. B: Schematic of the Cd9 protein with location of premature stop codon for Cd9a (red arrow) and Cd9b (green arrow) at a predicted 70 aa and 36 aa respectively. EC1/2 = Extracellular domain 1/2. C-D: Sequence chromatograms of genomic DNA from (c) WT and (d) cd9a^la61 allele. Deleted base pairs are underlined in red and inserted base pairs in blue. E-F: claudin b expression in WT at (e) 30 hpf and (f) 48 hpf. G-H: claudin b expression in cd9 dKO mutants at (g) 30 hpf and (h) 48 hpf. I-J: Graphs showing pLLP measurements in WT and cd9 dKOs; (i) migration of the cd9 dKO primordium at 36 hpf is significantly delayed compared to WT. (j) There is no significance in number of neuromasts deposited. Significance was assessed using an unpaired t test. N = minimum 9, p = <0.001, n1 = neuromast 1 etc. Bars show mean +/- SD.

Acknowledgments

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