Fig. S2

Morpholino specificity demonstrated by knockdown of GFP reporter expression or transcript mis-splicing in morpholino-treated embryos. Single cell zebrafish embryos were injected with morpholino, rhodamine dextran, and a GFP reporter plasmid and assayed for GFP expression 24 h later. (A and B) Bright-field images of 24 hpf embryos injected with control morpholino (C-MO) (A) or SOCS3a targeting morpholino (B) plus SOCS3a-GFP reporter plasmid. (C and D) Red fluorescence from rhodamine dextran demonstrating efficient injection in both embryos. (E and F) GFP expression is present in many cells of the C-MO-injected embryo (arrows in E) while GFP is absent in the target morpholino injected embryo (F). Auto-fluorescence present in the yolk is denoted by an asterisk. (G) Tabular results of embryo counts for each reporter and morpholino used in this study. In all cases, the C-MO had little effect on reporter expression while each targeting morpholino significantly decreased the number of GFP-positive embryos. (H) Single cell embryos were injected with control (C) or KLF splice-site targeted (SP) morpholino and RNA isolated 24 h later for analysis of mis-splicing using RT-PCR. Note that the splice site targeted morpholinos resulted in mis-splicing of each gene with mis-splicing of KLF7a being complete and mis-splicing of KLF6a being partial.