Identification and expression of TsATG4B. A qPCR analysis of TsATG4B gene transcription levels at various stages of T. spiralis development. 18S rRNA was used as the housekeeping gene control. ***P < 0.001, compared to the level in ML. B Results of TsATG4B analysis by SDS-PAGE. Lane M: protein molecular weight marker; lane 1: somatic proteins of T. spiralis ML; lane 2: ES proteins of ML; lane 3: purified rTsATG4B protein product; C Analysis of rTsATG4B antigenicity by Western blotting. Lanes 1, 4, and 7: soluble proteins from ML; lanes 2, 5, and 8: ES proteins of ML; lanes 3, 6, and 9: purified rTsATG4B. Proteins were probed with infection serum (1:100 dilution, lanes 1, 2, and 3), anti-TsATG4B serum (1:100 dilution, lanes 4, 5, and 6) and normal mouse serum (1:100 dilution, lanes 7, 8, and 9). D Native TsATG4B in soluble proteins of different developmental stages of T. spiralis (lane 1: NBL; lane 2: ML; lane 3: IIL; lane 4: 3 days AW; lane 5: 6 days AW) was recognized by anti-TsATG4B serum.