Figure 2.

Zebrafish develop antibodies against tick α-Gal and proteins. (A) The IgM antibody titers against α-Gal were determined by ELISA, represented as the average ± SD OD at 450 nm and compared between fish treated with saliva, α-Gal, PGE₂, or α-Gal + PGE₂, and the PBS-treated control group by Student t-test with unequal variance (^*p < 0.005; N = 7–9). (B) The IgM antibody titers against tick salivary gland proteins were determined by ELISA, represented as the average ± SD OD at 450 nm and compared between fish treated with saliva, α-Gal, PGE₂, or α-Gal + PGE₂ and the PBS-treated control group by Student t-test with unequal variance (^*p < 0.001; N = 7–9). (C) The α-Gal levels were determined by ELISA in salivary glands from unfed and partially fed ticks and saliva from fed ticks in comparison with pork kidney (α-Gal positive) and human HL60 cells (α-Gal negative) as positive and negative controls, respectively. The results were converted to α-Gal content per sample using a calibration curve (R² = 0.992; Supplementary Figure 5A) and compared between all samples and negative (lines) or positive (^*p < 1E-8) controls by Student t-test with unequal variance (p < 0.05, N = 3 biological replicates).