Fig. S3

rhouaATG morpholino (MO) represses GFP expression of arhoua-GFPreporter construct andrhouasplice MO perturbs splicing of therhouagene. (A-D) rhoua ATG MO1 blocks GFP expression from a construct in which the target sequence is placed upstream of the GFP coding sequence (rhoua-ATG-GFP). (A, B) RNA injected from the rhoua-ATG-GFP construct results in GFP expression throughout the embryo (n = 43/43 GFP+). (C, D) Injection of the rhoua ATG MO represses the expression of GFP due to this construct (n = 0/57 GFP+). (A, C) Brightfield microscopy. (B, D) GFP fluorescence. (E) Gel electrophoresis image shows that the rhoua splice MO2 blocks the proper splicing of rhoua and results in five different alternative splice products (Band 1-5). (F) Diagram of the rhoua genomic structure shows how the five different rhoua alternative splice products were produced by the rhoua splice MO2 (green). Light blue boxes represent exons. Purple lines represent new splice isoforms created due to the splice morpholino. Red line represents the normal splicing between exon 1 and exon 2.