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Fig. 1 fstl1a and fstl1b knockdown affect primary cilia formation in early embryos. a) Diagram depicting the partial genomic structure of fstl1a and fslt1b to show the site of morpholino binding (grey bar) to mRNA and the location of primers (arrows) used to evaluate knockdown efficiency. Also, schematic representations of the corresponding protein structures are shown. b) Gel image showing RT-PCR results demonstrating impaired amplification of fstl1a and fstl1b in the MOfstl1a and MOfstl1b morphant embryos, respectively. Gapdh was used as control. c) Low magnification images of 48 hpf wild-type and morphant embryos, and graphs showing the distribution of embryos in distinct phenotypic classes according to severity at the morpholino doses tested. The pooled data from three experiments are presented; the number of embryos analysed in each condition is indicated within brackets. d) Dot plot with the quantification of ciliary length in Kupffer vesicle of 12 somite-stage control and morphant embryos. The pooled data from a single experiment are presented; the total number of cilia measured/number of embryos is indicated in brackets. Plot: individual cilia length values and median ± CI (95%) (*) p < 0.0001; (**) p = 0.0019; Games?Howell test. e) Representative images showing primary cilia (evidenced through acetylated tubulin immunostaining) in the nasal pit and otic vesicle of 48 hpf embryos injected with either 6 ng of control MO, MOfstl1a or MOfstl1b. Phalloidin staining of F-actin was used to delineate the organ. Scale bars: C: 1 mm; E: 10 µm.