XB-IMG-175397
Xenbase Image ID: 175397
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Supplementary figure 3:The phenotype obtained after knock down of P-TEFb components. (A) Graph showing the percentage of embryos displaying a pigment loss phenotype after injection of 100ng standard control morpholino n=119, 60ng CYCLINT1 morpholino n=95, 100ng CDK9.L morpholino n=62 and a combination of both 40ng CDK9.S and 60ng CDK9.L Morpholinos n=73. *=P<0.05 by Kruskall Wallis statistical test. (B) Embryos were morpholino injected in 1 cell of a 2 cell stage embryo left to develop to stage 45 then underwent alcian blue stain to observe craniofacial cartilage. (i) 100ng control morpholino showing wild type craniofacial cartilage. (ii) Schematic of normal wild type craniofacial cartilage M=Meckels cartilage, C=Ceratohyal, B=Branchial cartilage. (iii) Injection of 100ng CDK9.S morpholino shows abnormal development of the branchial cartilage (black arrow head). (iv) Injection of 60ng CYCLINT1 morpholino shows abnormal development of the branchial cartilage (black arrow head). (C) Sensory neuron function was assayed using a poke and stroke assay to assess touch response. This response was scored as follows; swim=1, twitch=0.5, no response=0. Embryos were injected in both cells of a 2 cell stage embryo with either 100ng of a control morpholino, 100ng Cdk9.S morpholino or 60ng CyclinT1 morpholino. Poke: CoMo 9.0 ± 0.25 (n=30), Cdk9.S Mo 1.83 ± 0.14 (n=30), CyclinT1 Mo 1.66 ± 0.52 (n=30). Stroke: CoMo 9.83 ± 0.14 (n=30), Cdk9.S Mo 1.83 ± 0.52 (n=30), CyclinT1 Mo 2.33 ± 0.38 (n=30) *=p<0.05 by one-way ANOVA versus standard control morpholino. CoMo = control morpholino. (D) Graphs showing the percentage of embryos displaying an expansion, wild type, partial loss or loss of expression pattern by in situ hybridisation after injection of 60ng CYCLINT1 morpholino (top panel), 100ng CDK9.S morpholino (middle panel) and a standard control morpholino (bottom panel). Como injected c-myc n=27, sox10 n=29, slug n=38, foxd3 n=35, sox9 n=33, zic1 n=31, zic3 n=55, pax3 n=40, sox2 n=27. Cdk9.S mo injected c-myc n=32, sox10 n=51, slug n=51, foxd3 n=55, sox9 n=31, zic1 n=32, zic3 n=62, pax3 n=38, sox2 n=44. CyclinT1 injected c-myc n=30, sox10 n=46, slug n=37, foxd3 n=32, sox9 n=39, zic1 n=32, zic3 n=70, pax3 n=54, sox2 n=54. Image published in: Hatch VL et al. (2016) Copyright © 2016. Image reproduced with permission of the Publisher, Elsevier B. V. Larger Image Printer Friendly View |