Fig. 3

New annotations for Olfr1507 and Olfr15 3’UTR isoforms and experimental validation. a RNA-Seq raw data (see Fig. 1B for legend). b 3’UTRs generated using our own Cufflinks analysis. c 3’UTRs identified using our dedicated workflow applied to the male_IS2014 dataset with the STAR and IsoSCM algorithms. S: short, M: medium, L: long; XL: extra-long. Black vertical box: CDS end; black horizontal bar: 3’UTR. d Relative abundance of the alternative 3’UTR isoforms annotated in silico using our IsoSCM pipeline. e In silico validation of the 3’UTRs annotated using our IsoSCM pipeline by the presence of canonical PASs. Vertical bar = predicted canonical (AAUAAA or AUUAAA) PAS. f Experimental validation by RL-PAT (RNA Ligation mediated PolyAdenylation Test, Table 1). ✔ = genuine polyA site. *: XL site for Olfr1507, M1 and L sites for Olfr15 could not be tested by RL-PAT due to repeat sequences upstream of the polyA sites. g and h Experimental validation by northern blot. Total OM RNAs were separated on agarose/formaldehyde gels and transferred onto nitrocellulose membranes. The presence of Olfr mRNAs was detected following hybridization with DIG-labeled antisense probes either in the CDS region (CDS probe) or between the 3′S and 3’M ends (3’M probe; see Table 3 for detailed probe description). g The major isoform of the Olfr1507 mRNA shows a short 3’UTR (≈3-kb dark band with CDS probe, not detected with 3’M probe); additional isoforms having longer 3’UTRs are present at low abundance (4.7 to 6.9-kb faint bands with both probes); ♯ highest band (> 7 kb) corresponds to an intron-retaining transcript for Olfr1507 (Additional File 4: Fig. S2). h Olfr15 shows 2 major isoforms, the first one with a short 3’UTR (≈2.5-kb band with CDS probe, not detected with 3’M probe = 3′S), and the second one with a longer 3’UTR (≈4.5-kb band with both probes = 3’M1/2).