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Fig. 3. ALK2 is required for the expansion of the organizer domain. a The efficacy of the ALK2MO was determined by western blot analysis. RNA encoding a myc-tagged version of the ALK2 protein with the ALK2MO recognition sequence was co-injected into embryos with increasing amounts of ALK2MO. Early gastrula protein extracts were subjected to electrophoresis and immunodetection with the 9E10 anti-myc monoclonal antibody. To control the loading and transfer steps, the blot was stripped and re-probed with an anti-actin antibody. b–m Embryos were injected with an antisense morpholino oligonucleotide specific for ALK2 (ALK2MO; 400 pg/embryo) or control MO (coMO). At the onset of gastrulation, embryos were analyzed by in situ hybridization with chordin (b–e), gsc (f–i), and ADMP (j–m) specific probes to determine the effect on their expression domain. The domain of expression was determined by measuring the arc in degrees and calculating the relative size compared to control embryos. The relative (%) arc of the expression domain was determined. Chordin analysis: control, n = 13; ALK2MO, n = 12; coMO, n = 13, gsc analysis: control, n = 11; ALK2MO, n = 12; coMO, n = 10, and ADMP analysis: control, n = 9; ALK2MO, n = 10; coMO, n = 8. p values were calculated compared to controls. Statistical test; Dunnett’s (ANOVA) multiple comparisons test. *p < 0.05, **p < 0.01, ns not significant Image published in: Leibovich A et al. (2018) © Fainsod et al. This image is reproduced with permission of the journal and the copyright holder. This is an open-access article distributed under the terms of the Creative Commons Attribution license
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