Supplementary Figure 7: Characterization of a second antisense morpholino oligomer targeting Pkd1 mRNA splicing (Pkd1-sMO2).

Xenopus embryos were injected with 3.2 pMol Pkd1-sMO2 into all blastomeres at the 2- to 4-cell stage and cultured until sibling control embryos reached stage 35, 40, or 43. (a–c) 3D reconstruction of pronephric kidneys by immunofluorescence staining using the 3G8 and 4A6 antibodies at stage 40. Representative images are shown in panels (a,b) and results are summarized in (c). (d) Quantification of embryos displaying edema at stage 43. (e) RT-PCR of Pkd1 mRNA in uninjected and Pkd1s-MO2-injected embryos at stage 35. PCR products representing correctly spliced mRNA (200 bp) in uninjected embryos or Pkd1 mRNA retaining the intron (850 bp) in Pkd1-sMO2-injected embryos are indicated. All experiments were performed using three independent biological replicates. The number of embryos analyzed is indicated above the bars in (c,d).