Fig. 1.

Fig. 1.

cdkl5 expression and validation of the cdkl5^sa21938 zebrafish mutant strain. (A) Schematic of the zebrafish Cdkl5 long (Cdkl5-L) and short (Cdkl5-S) protein isoforms showing the location of the nonsense (PTC) mutation and primers used for RT-PCR analyses. The long (3936 bp) and short isoforms (3479 bp) both contain exons 1-15 and exons 17-18 (grey), and differ by the inclusion of exon 16 (white) and the lengths of their 5′ untranslated region (UTR) (orange) and 3′ UTR (green) regions. (B) RT-PCR analyses of cdkl5-L and cdkl5-S transcripts from 1-6 dpf.  β-Act was amplified as a positive control. (C) cdkl5 mutants contain a C>A change in exon 11 of the gene (amino acid 528 out of 1039 and 1080 of the short and long isoforms, respectively), creating a premature termination codon (PTC), as seen in the sequencing chromatograms from cdkl5^+/− fish. (D) qRT-PCR analysis demonstrated that cdkl5-L and total cdkl5 mRNA levels are significantly reduced in cdkl5^−/− embryos compared to cdkl5^+/+ siblings at 3 dpf. Data are mean±s.e.m. for three independent experiments (with 20 pooled fish per experiment), *P<0.05 (one-way ANOVA).