Fig. 4

Expression of TsTryp at different developmental stages of T. spiralis. a qPCR test for TsTryp transcription level at different T. spiralis stages, calculated based on the 2^−ΔΔCt method. The fold changes in TsTryp gene level were normalized to the respective GAPDH control. b Western blot of TsTryp protein expression level during various worm stages. TsTryp expression in crude antigens of ML larvae, IIL, 3-/6 dpi AW and NBL was assayed by western blot using immune serum against rTsTryp. The data were derived from three separate tests. Asterisk indicates a statistical significant difference at *P < 0.05 compared to the ML stage. AW, Adult worms; d, day post-infection; ES, excretory/secretory; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; h, hour post-infection; IIL, intestinal infective larvae; ML, muscle larvae; mRNA, messenger RNA; NBL, newborn larvae; qPCR, quantitative PCR; rTsTryp, recombinant T. spiralis trypsin