Fig. 2
From: Knockdown of myorg leads to brain calcification in zebrafish
myorg knockdown showed brain calcifications in zebrafish. A Schematic depiction of the myorg transcript after E2I2-MO (a) and ATG-MO (b) administration. B The effectiveness of myorg knockdown was confirmed by RT-PCR after morpholino injection of zebrafish larvae at 2 dpf. Injection of 4 ng of myorg morpholino altered the splicing between Exon 2 and Intron 2, as revealed by a shift in PCR bands between control (619 bp) and myorg morpholino injected embryos (137 bp). C Sanger sequencing confirmed the transcription of skipping Exon 2 after the injection of myorg-E2I2-MO. dpf, days post-fertilization. D Dorsal views of zebrafish embryos injected with control morpholino oligonucleotides (Control-MO), myorg-E2I2-MO, and myorg-ATG-MO. MO injected embryos were stained with calcein at 3 dpf. Compared to Control-MO (a–c), myorg-E2I2-MO (d–f), and myorg-ATG-MO (g–i) showed potent brain calcifications as green fluorescence in the zebrafish brain. (a, d, g) bright field; (b, e, h) calcein staining; the magnified zooms of the red dashed box are shown in c, f, i. E Quantification of the brain calcifications in the myorg-E2I2-MO and myorg-ATG-MO injected zebrafish at 3 dpf compared to that in Control-MO injected zebrafish. F Time-course analysis of brain calcifications in the myorg-E2I2-MO knockdown zebrafish. Means ± SEM, n = 10, Student’s t-test, ****P < 0.0001, ***P < 0.001, **P < 0.01. Scale bar, 50 μm. dpf days post-fertilization