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, Embryos were injected at the 1-cell stage with 1 pmol MOex5, 200 pg map9-YFP RNA or both. Map9-YFP-injected embryos developed normally up to mid-epiboly and later stages. 40% of MOex5 morphants showed defects at mid-epiboly and 20% were blocked at the sphere stage. Following co-injection of MOex5 and map9-YFP RNA, about 80% of embryos developed normally up to mid-epiboly, but died at ~9-10 hpf. (B) Representative images of embryos injected with map9-YFP RNA (at mid-epiboly, n=102), with MOex5 (showing developmental delay and defects before mid-epiboly, n=85) or with both MOex5 and map9-YFP RNA, Map9-YFP RNA can partially rescue the phenotypes of MOex5 morphants. (A)
, Morpholino-mediated depletion of map9 leads to increased apoptosis. (A-D) Zebrafish embryos were injected, or not (control), at the 1-cell stage with 1 pmol of MOex5 for observation at 8 hpf
, TUNEL-positive cells (green) were counted in injected (B, D) and control embryos (A, C) (n=10/assay). (E) Left panel, number of TUNEL-positive cells in the control and MOex5-injected embryos depicted in A-D, n= 5 embryos per assay
Number of TUNEL-positive cells in MOex5-and MOex5 + MOtp53-injected embryos (n=20 embryos per assay, total cell count ~564). (F) Map9 depletion does not induce tp53 expression. Embryos were injected at the 1-cell stage with 1 pmol of MOex5. RNA from 7 hpf map9 morphants and control embryos was used to quantify tp53 gene expression by RT-qPCR. Expression data were normalized to ?-actin. Experiments were made in quadruplicate, p.50 ,
, MOex5 morphants at 7 hpf, and n=39/44 embryos at 24 hpf. (G) Co-injection of MOex5 (1 pmol) and
5 pmol) does not rescue MOex5 effects. Embryo mortality was not rescued after tp53 depletion by morpholino. Embryo mortality after injection of a control MO (MOmis) was not significantly different compared to non-injected embryos. This experiment was made in duplicate ,