The Journal of Neuroscience, December 20, 2006, ():
Netrin/DCC Signaling Controls Contralateral Dendrites of Octavolateralis Efferent Neurons
J. Neurosci. Suli et al.
26: 13328
Supplemental Data
Files in this Data Supplement:
- supplemental material
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Supplemental Figure 1. dcc is misspliced after injection of DCC-MO3. a) DCC-MO3 is designed to bind the intron1/exon1 splice donor. (P1-P3, RT-PCR primers.) b) RT-PCR on 36 hpf DCC-MO3 morphants shows a decrease in properly spliced dcc mRNA and a new, larger product (arrows). P1/P3, pre-mRNA; P1/P2, spliced mRNA; beta-actin, loading control. c) Sequencing this larger band shows utilization of a cryptic splice donor within intron1, leading to an insertion of 72 bp (24 aa) after exon1 in the dcc mRNA. d) Schematic showing missplicing of dcc after injection of DCC-MO3.
- supplemental material
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Supplemental Figure 2. netrin1a mRNA is greatly reduced after injection of Netrin1a splice-blocking MO (Netrin1a-SBMO). a) The Netrin1a-SBMO is designed to bind the intron1/exon1 splice donor. (Genomic structure not to scale. F1, R1, R2: RT-PCR primers.) b) RT-PCR on 36 hpf netrin1a/netrin1b morphants using primer pairs that amplify unspliced netrin1a pre-mRNA (F1+R1); properly spliced netrin1a mRNA (F1+R2); and beta-actin mRNA as a loading control (beta-actin). Normal mRNA seems to be completely abolished by injection of the Netrin1a-MO, while unspliced pre-mRNA levels are increased. Reactions lacking reverse transcriptase (-RT) confirm that amplified bands reflect RNA and not genomic DNA.
