|
Fig. 2 Chp function is required for epibolic movements.
(A) Positions and sequence of Chp morpholino-oligoncleotides used in the study. The Chp MO1 and MO2 (corresponding to the reverse strand) are represented in purple and green respectively; lowercase sequence is 5′ UTR region. (B) Phenotypic examination of Chp morphants at 8 hpf revealed delayed closure of the yolk plug when compared with the un-injected controls. Rescue of the phenotype was performed by co-injecting Chp MO 1/2 with WT Chp (Zf) mRNA. Co-injection of 25 pg Chp mRNA with 5 ng Chp MO1/2 efficiently reduced the number of yolk plug defects at 8 hpf. Phenotypic analysis shows significant rescue in the delay of yolk plug closure at 8 hpf. (C) Immuno-histochemical staining of un-injected controls and Chp morphants at 60% epiboly using Chp antibody raised and purified from two animals (Pab1 and Pab2). Confocal images show endogenous Chp is strongly depleted Chp morphants. Decreased protein levels were observed in EVL, DEL and YSL layers with Chp MO. Embryos were counter-stained with DAPI. Scale bars = 20 μm.