Additional file 1: Table S1. of Characterization of a novel aspartyl protease inhibitor from Haemonchus contortus

Primers for amplification of API gene. Table S2. Primers for real time PCR. Table S3. Primers for quantification of cytokines transcription. (DOCX 18 kb

Additional file 3: Figure S1. of Characterization of a novel aspartyl protease inhibitor from Haemonchus contortus

Amplification efficiencies of target gene (API) and endogenous reference (β-Tubulin) gene were verified to be similar by real time PCR. Figure S2. Amplification efficiencies of target genes (IL-2, IL-4, IL-10, Il-17, IFN-γ and TGF-β) and endogenous reference (β-Actin) gene were verified to be similar by real time PCR. (DOCX 57 kb

Additional file 1: Figure S1. of Arginine kinase from Haemonchus contortus decreased the proliferation and increased the apoptosis of goat PBMCs in vitro

Cloning and expression of Hc-AK gene. The recombinant plasmid pMD19T-AK (a) and expression plasmid pET32a (+)-AK (b), were verified by restriction digestion with BamH I and EcoR I. Figure S2. Multiple sequence alignment of Hc-AK. Amino acid sequence of Hc-AK with that from other species, H. contortus (CDJ90032), C. briggsae (XP_002645008), C. brenneri (EGT52941), C. elegans (NP_509217), H. glycines (AAO49799), O. dentatum (KHJ89945), A. duodenale (KIH65495), D. viviparus (KJH41917), V. pacos (XP_006219889) and N. americanus (XP_013303820) using CLUSTAL W method and GeneDoc ( http://www.psc.edu/biomed/genedoc/ ). Figure S3. Phylogenetic analysis for Hc-AK gene. A phylogenetic tree was constructed by neighbour -joining method to verify relationships between the amino acid of Hc-AK to that of other nematode species, using MEGA ver. 6.1 programme. Figure S4. N-terminal signal peptide prediction. The amino acid sequences of Hc-AK (NCBI accession numbers: JX422018.1 was used to predict N-terminal signal peptides by SignalP 4.1 Server. Figure S5. Membrane protein prediction by using TMHMM Server v.2.0. The amino acid sequences of Hc-AK (NCBI accession numbers: JX422018.1) was analysed to predict transmembrane structures using TMHMM Server v.2.0. There were no transmembrane domains predicted in this protein structure. http://www.cbs.dtu.dk/services/TMHMM/ . Figure S6. Prediction of B and T cell epitopes. Protein sequence of Hc-AK (NCBI accession numbers: JX422018.1) was used for the prediction of the B cell and T cell epitopes, that revealed 16 peptides of B cell epitopes and 17 T cell epitopes. (DOCX 736 kb