Figure 4

qRT-PCR analysis of midgut antimicrobial peptide (AMP) expression in female Culicoides sonorensis . Female midges were fed blood or sucrose and midguts (n = 15 per time-point) were dissected at 3, 8, 12, and 24 h after feeding. Relative AMP expression was determined using the methods of Pfaffl [31] for attacin m.3140 (A), attacin m.7821 (B), defensin m.9998 (C), defensin m.9997 (D), and cecropin m.10000 (E), with teneral midges serving as the calibrator condition and incorporating the reference gene EF1b. Error bars represent SEM taken from three biological replicates. Asterisks denote change in expression from baseline (teneral) levels (P < 0.05). n.d., not determined (i.e., threshold cycle not crossed within 40 cycles). The details of each of the three biological replicates, including P-values, are available in Additional file 2.