Fig. 2

Morpholinos Targeting the Precursor miRNA Interfere with miRNA Maturation (A) Design of morpholinos targeting the precursor miRNA. (B) Northern blot analysis of miR-205 in 30-h-old embryos injected with different morpholinos against pri-miR-205. 5S RNA serves as a loading control. (C) Time series of miR-205 expression after injection of mature, no lap loop, and drosha star morpholinos against pri-miR-205. (D) Northern blot analysis of miR-205 derived from embryos injected with a GFP-pri-miR-205 transcript and four different morpholinos targeting pri-miR-205. Co-injected miR-206 serves as an injection and loading control. Embryos were collected 8 h after injection. (E) GFP expression in 24-h embryos injected with morpholinos and a GFP-pri-miR-205 construct as used in (C). Pri-miR-205 is positioned just upstream of the polyA signal in the 3′ UTR of the GFP mRNA. Red fluorescent protein (RFP) serves as an injection control. (F) RT-PCR analysis of injected GFP-pri-miR-205 mRNA with (+) and without (-) co-injected morpholinos. Luciferase serves a an injection control. Embryos were collected 8 h after injection. (G) Northern analysis of the effect of morpholinos on an injected miR-205 precursor. Embryos were collected 8 h after injection. WT, wild type.