Figure 5. Relative expression of target genes ACT, UBQ, and NMHC in relation to LM29173 load.

(A–C) Relative expression of target genes β-actin (A; ACT), (B; UBQ) and (C; NMHC) in relation to expression of reference gene elongation factor-1α (EF1A) in specimens from two haplotype clusters (northern and southern) with above (grey) and below (white) average LM29173 copy number (±1 SE). (D–F) Correlation between viral load and relative expression of ACT (D), UBQ (E), and NMHC (F) in relation to EF1A reference gene expression. Quantities of target amplicons were standardized by reference gene EF1A using the following equation: (Target_RT–Target_NRT)/(EF1A_RT − EF1A_NRT), where RT and NRT indicate samples that have been reverse transcribed via Superscript III (Invitrogen, Carlsbad, CA, USA), or not reverse transcribed (no-RT control), respectively. (G) Gene expression (reads per kilobase of transcript per million mapped reads; RPKM) of ACT, NMHC, and UBQ per transcriptome library in each of three stations: Lake Superior station 066 (SU066), Lake Michigan station 40 (Mi40) and Lake Michigan station 27 (Mi27). Libraries are ranked from left to right by increasing LM29173 load.