QPCR of selected transcripts with immune relevant functional annotations identified in the reverse liver SSH library (designed to be enriched for transcipts down-regulated by heat-shock). The RQs (relative quantities), normalized to 18S ribosomal RNA expression and calibrated to the individual with the lowest expression of each gene of interest (see methods section), are presented as averages ± SEM. The levels of gene expression of TLR22 (Toll-like receptor 22: A), nuclear protein 1 (NUPR1: B), alpha 1-microglobulin/bikunin (Bikunin: C) and immunoglobulin heavy chain, secretory form (IgM: D) are shown for the control transferred (CT) and heat-shocked (HS) groups before heat-shock (BHS), at the cessation of heat-shock (CS), 3h after the cessation of heat-shock (3ACS), and 12h after the cessation of heat-shock (12ACS). Different letters indicate significant differences between sampling points within the same treatment (p< 0.05). * indicates significant differences between CT and HS groups within a given sampling point (p< 0.05). Numbers in the boxes represent overall fold-changes. For each treatment, overall fold up-regulation was calculated relative to the appropriate before heat-shock (BHS) values as (average RQ for a time point)/(average RQ of the appropriate group at the BHS time point), and overall fold down-regulation was calculated when necessary (i.e. if overall fold upregulation was < 1) as the inverse of overall fold up-regulation. Each panel shows the expression for a given gene of interest in both tissues studied. L: liver. HK: head kidney.