Knock-Down of Cathepsin D Affects the Retinal Pigment Epithelium, Impairs Swim-Bladder Ontogenesis and Causes Premature Death in Zebrafish
Figure 4
Zebrafish cathepsin D down-regulation by two different morpholinos.
A) Schematic representation of S-MPO and T-MPO target regions in the CD RNA. The sequences of both MPOs are shown. B) Cloning of the 622bp, comprehensive of the S-MPO complementary site, from the genomic DNA of 4dpf larvae. C) Portion of the sequence of the 622bp region corresponding to the S-MPO complementary site. This experiment was performed two times with identical results. D) Western blotting of CD in homogenates of 4 dpf larvae deriving from control injection (CTRL), injection with splicing morpholino (S-MPO) and injection with translation morpholino (T-MPO). The filter was incubated with polyclonal antibody against rat CD, stripped and re-probed with monoclonal antibody against β-actin. The densitometry ratio of CD vs actin expression calculated from 5 independent experiments in double is shown. (a.u. = arbitrary units).