Fig. 4

Fig. 4

(a) Schematic diagram of the design of the exon skipping MO used in the study. (b) Schema representing the zebrafish cdon mRNA (zcdon) aligned with the corresponding Cdon protein indicating the domains targeted by cdon^spl8, cdon^spl11a, cdon^spl11d and cdon^spl14 MOs. The exons that encode the 52 and 32 UTR regions are depicted in blue; those that, when skipped, generate a translational frame shift are indicated in red; whereas those that, when skipped, maintain the reading frame are indicated in green. (c,f,i) RT–PCR analysis of the exon skipping function of cdon^spl8, cdon^spl14 and cdon^spl11a/cdon^spl11d MOs. For detailed information about the resulting bands, noted 1–9, please refer to Supplementary Fig. 8. (d,e,g,h,j,k) ISH analysis of pax2.1 expression pattern in cdon^spl8 (d,e), cdon^spl14(g,h) and cdon^spl11a/cdon^spl11d (j,k) MO injected embryos at 26–28 h.p.f. Pax2.1 expression domain is expanded in cdon^spl8 (d,e) and cdon^spl14 morphants (g,h) in comparison with their respective controls, whereas there was no difference in the pax2.1 expression domain of control and cdon^spl11a/cdon^spl11d morphants (j,k). (l) Quantification of the optic stalk pax2.1-positive expression domain in embryos injected with the different MOs at 26–28 h.p.f. (***P< 0.001; Student’s t-test). The number of embryos analysed in each case is indicated in each column and are as follows: Cdon^ATG MO injection (control, n=28; MO, n=45); Cdon^spl8 MO injection (control, n=12; MO, n=44); Cdon^spl11a and Cdon^spl11d MOs injection (control, n=26; MO, n=65); Cdon^spl14 MO injection (control, n=22; MO, n=57). Error bars represent s.e.m. Scale bars, 100 μm. MWM, molecular weight marker.