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The phylogenies of the genes families with duplicated members show that the duplication timing dates back to the origin of the teleosts (Additional File 4, Figure S4).
Although the nature of the assay did not allow us to handle large number of cells, at least ten SPBs were analyzed for each time point to capture a trend for duplication timing.
Firstly, by determining the duplication timing through phylogenetics, and secondly by investigating the SCD gene loci in available tetrapod genomes representing various lineages.
Overall, the majority of genes flanking human ACSL1, ACSL5 and ACSL6 revealed conserved macrosynteny and therefore support the hypothesis that these regions are related, with the duplication timing coinciding with 2R.
The analysis of the flanking gene families revealed that those which are multi-copy and whose duplication timing coincides with vertebrate emergence, typically have their members localizing to Hsa4, Hsa5, Hsa8, Hsa10 and/or Hsa2.
Gene families with multiple members (e.g., oxysterol binding protein - OSBP) flanking these genes in humans had their phylogenetic history determined to clarify if the duplication timing coincided with 2R (not shown).
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Phylogenetic analysis was then performed when no previous study was available to determine duplication timings (if duplication had occurred).
Through phylogenetics we were able to reconstruct the Acsl gene duplication timings in relation to the divergence of major vertebrate groups.
This strategy aimed at defining the presence/absence of invertebrate orthologues and the duplication timings of the selected gene families (both these questions are key predictions of paralogy regions).
Correlations between gene copy numbers and genome size as well as whole-genome duplication events timing was estimated using the Pearson correlation with a two tailed significance test.
If these three subtypes are products of the so-called two-round whole genome duplications, their timing is consistent with an emerging hypothesis that the common ancestor of all extant vertebrates already experienced these events [ 24, 25].
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