Multiple alignment analysis of EtPDIL of <i>Eimera tenella</i> with PDIL from other apicomplexan parasites.

<p>Shown are sequences from <i>Plasmodium cynomolgi</i> (XP_004221713), <i>Plasmodium vivax</i> (XP_001614725), <i>Eimeria mitis</i> (CDJ34317.1). Deduced protein sequences were used in the Clustal W sequence alignment program. Asterisks, identical amino acids.</p

Quantitative real-time RT-PCR of EtPDIL expression in <i>E. tenella</i> developmental stages.

<p>UO, unsporulated oocysts; SO, sporulated oocysts; Spz, sporozoites; Mrz, merozoites. Bars not sharing the same letters were significantly different (P<0.05).</p

Inhibition of sporozoite invasion <i>in vitro</i> by antibody against rEtPDIL.

<p>Anti- rEtPDIL, rabbit antiserum against recombinant EtPDIL protein; NA, naive rabbit serum. All assays were performed in triplicate. ** P<0.01 for differences between treatment with antibody against rEtPDIL and naïve rabbit serum at the same IgG concentration.</p

Western blots.

<p>(A) Purified recombinant EtPDIL (diaminobenzidine as substrate). Rabbit sera against sporulated oocysts of <i>E. tenella</i> or anti-GST monoclonal antibody was used as primary antibody. Lane 1, protein marker. Lane 2, anti-GST monoclonal. Lane 3, antisporulated-oocysts serum. Lane 4, naive rabbit serum. (B) Protein lysates from four different life cycle stages of <i>E. tenella.</i> Rabbit sera against rEtPDIL or mouse monoclonal anti-α-tubulin was used as primary antibody. Spz, sporozoites; SO, sporulated oocysts; UO, unsporulated oocysts; Mrz, merozoites.</p

Localization of EtPDIL in different stages of <i>E. tenella</i> by indirect immuofluorescence using rEtPDIL antibody.

<p>(A) Sporozoites (Spz) in PBS; (B) Spz in complete medium. Infected DF-1 cells were collected at indicated hours post infection (p.i.). (C) 2 h p.i., intracellular sporozoites (iSpz); (D) 12 h p.i., intracellular sporozoites (iSpz); (E) 24 h p.i., trophozoites (Tropho); (F) 48 h p.i., immature schizonts (iSc); (G) 60 h p.i., immature schizonts (iSc); (H) 68 h p.i., immature schizonts (iSc); (I) 72 h p.i., mature schizonts (mSc); (J) 85 h p.i., intracellular merozoites (iMrz); (K) Merozoites (Mrz) in PBS.</p

Full cDNA and deduced amino acid sequence of EtPDIL.

<p>Underlined, start and stop codons; double underlined, CAAT box in the 5′-UTR; gray, non-classical thioredoxin-like motif (SXXC); wavy underlined, N-myristoylation site.</p

Expression of recombinant EtPDIL in <i>Escherichia coli</i> by SDS-PAGE.

<p>Lane 1, protein marker; lane 2, IPTG-induced recombinant EtPDIL protein at 0 h; lane 3, IPTG-induced recombinant EtPDIL protein at 6 h; lane 4, purified recombinant EtPDIL.</p

Primer sequence and name.

<p>Primer sequence and name.</p