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Fig. SX.5
spp1 gene knockdown in zebrafish.
(a) Five dpf zebrafish embryos stained with alcian blue for cartilage. Wild type, ATG morphant and E2-I2 morphant embryos are shown. Morphant embryos do not show any discernible change in cartilage matrix formation. (b) E2-I2 MO inhibits splicing of spp1 in zebrafish. Upper panel shows the position of RT-PCR primers (P1, P2 and P3) in relation to spp1 gene. Lower panel shows the RT-PCR fragments separated on a 3% agarose gel. The wild type spliced product is represented by a 120 bp fragment, whereas unspliced products are represented by the 87 bp and 230 bp fragments. Splicing of intron 2 is considerably inhibited in 4-dpf and 5-dpf embryos injected with E2-I2 MO, while normal splicing occurs in 4-dpf and 5-dpf embryos that are wild-type or injected with E2-I2 control MO.