Notably, the radiation unresponsive transcript variants of Zfp365, Ano3 (Fig. 6) and Usp2 (not shown) showed deviating expression levels with either no change, or decreased expression during development or differentiation.
In all these studies fidelity, reproducibility and linearity of RNA amplification has been a major concern and increasingly refined statistics have been used accordingly (correlation, fold change, T-test, ANOVA; [ 23]) to identify amplification biases through deviating expression patterns between amplified and un-amplified material.
Among the 1,264 unique samples that were eventually selected there were two samples that we excluded manually because they showed deviating expression levels from what we expected for their presumed tissue and because they had barely passed various other filter criteria.
Consistent with characteristics of other takeout family members, deviate expression is responsive to photophase conditions (p < 0.1), and feeding, temperature, and JH (p < 0.05).
These findings are among the first linking the expression of a termite gene with eusocial behavior; they illustrate the connection between deviate expression and trailing behavior, which is a key evolutionary adaptation vital to subterranean social insects such as termites and ants.
Allowing for mismatches in this step augments this even more, leading to deviating expression profiles.
Of this gene, several exons showed a deviating expression profile after radiation, indicating the induction of unknown splice variants (Fig. 2E, left panel).
Although overall expression levels are similar between strains, a relatively high number of differentially expressed genes was due to deviating gene expression levels in BL6.
Only one cluster differing between line X and Y was associated with immunity, one could expect more because these lines differ in resistance against bacterial infection and showed deviating gene expression after an immune challenge [ 29– 35].
In Panel 1, the endogenous controls encoding RPL23a and EF1α were recognised thus confirming that some of the deviating expressions we observed on Fig. 6 were due to IVT induced over-expressions (see Fig. 2).
