Fig. 3
Temporal distribution of RhEcR and RhUSP isoforms in the salivary gland. Tissues were dissected from female ticks. Using qRT-PCR, relative RNA levels of RhEcR and RhUSP were measured at each time point. a In the period of blood-feeding, the expression of RhEcR2 was higher than RhEcR1 before E 24 h. After E 24 h, the expression of RhEcR1 was higher than RhEcR2 (ANOVA, RhEcR1/2/3, Engorged-48h vs Engorged-72h, F (2, 24), P < 0.0001; Engorged-72h vs Engorged-96h, F (2, 24), P = 0.0012; Engorged-96h vs Engorged-24h, F (2, 24), P < 0.0001; Engorged-24h vs Fed5, F (2, 24), P = 0.008; Fed5 vs Fed3, F (2, 24), P = 0.9877; Fed3 vs Engorged-0h, F (2, 24), P = 0.0018; Engorged-0h vs unfed, F (2, 24), P = 0.9877; RhEcR1, Engorged-48h vs Engorged-72h, F (2, 24), P = 0.1115; Engorged-72h vs Engorged-96h, F (2, 24), P = 0.9984; Engorged-96h vs Engorged-24h, F (2, 24), P < 0.0001; Engorged-24h vs Fed5, F (2, 24), P = 0.9877; Fed5 vs Fed3, F (2, 24), P > 0.9999; Fed3 vs Engorged-0h, F (2, 24), P > 0.9999; Engorged-0h vs unfed, F (2, 24), P = 0.9877; RhEcR2, Engorged-48h vs Engorged-72h, F (2, 24), P = 0.6662; Engorged-72h vs Engorged-96h, F (2, 24), P = 0.6662; Engorged-96h vs Engorged-24h, F (2, 24), P > 0.9999; Engorged-24h vs Fed5, F (2, 24), P > 0.9999; Fed5 vs Fed3, F (2, 24), P > 0.9999; Fed3 vs Engorged-0h, F (2, 24), P = 0.023; Engorged-0h vs unfed, F (2, 24), P = 0.9877). b The translation of RhEcR isoforms was detected with anti-mouse RhEcR polyclonal antibody. After E 24 h, the expression of all isoforms of RhEcR gradually decreased. c Two isoforms of RhUSP were detected during all time points, showing that the RNA levels of RhUSP1 were high and RhUSP3 were very low (ANOVA, RhUSP1, Engorged-48h vs Engorged-72h, F (1, 17), P < 0.0001; Engorged-72h vs Engorged-96h, F (1, 17), P < 0.0001; Engorged-96h vs unfed, F (1, 17), P < 0.0001; unfed vs Engorged-24h, F (1, 17), P = 0.9855; Engorged-24h vs Fed5, F (1, 17), P = 0.1282; Fed5 vs Fed3, F (1, 17), P = 0.9855; Fed3 vs Engorged-0h, F (1, 17), P > 0.9999). The data represent the mean ± SD of the experiment (10 ticks/time point) performed in triplicate and normalized to EF1α, and 2 way ANOVA multiple comparisions for statistical differences between multiple groups were used (α = 0.05)