Figure 1.

Design of the experiment and qPCR analysis. Successive steps are linked with gray arrows indicating procedures being done. The text above the arrows indicates the specific cytokine genes used for each step, and the text below the arrows indicates the treatment group(s) used for that step [control (C)]. The text inside the arrows indicates the sample size per tissue and the metric of gene expression used [relative expression (R) ratio (41) calculation and standardized expression quantity (stQ) (42, 43)]. The stQ quantification was used for analyses where statistical comparisons between treatments and controls were necessary, as this is not possible using the R ratio. Blue triangles indicate traits for which associations to cytokine expression were tested. For isolate differences in cytokine expression in Harderian gland and choana, only a subset of cytokines was examined, and thus those two tissues are in parentheses.