Fig 1.
Morphology of different life cycle stages of E. multilocularis.
A: Egg; B: Oncospheres (non-activated); C: Oncospheres (activating) with the hooks dispersive and body swelling; D: Oncospheres (activated) with the hooks aggregation in the smaller lobe after 24 hours activation; E: 4-week metacestodes miniature vesicles; F: Metacestodes small vesicles cultivated in vitro; Bar: 10μm (A-E), 1mm (F); Arrowhead: Miniature vesicles.
Table 1.
Primers for real-time PCR.
Table 2.
Overview of the sequencing reads.
Fig 2.
Real-time PCR validation of genes at kinds of expression level.
The y-axis indicates the value of relative expression level (2-ΔΔCt) by real-time PCR and log2Ratio of Nonc1/Cmet by Next-generation sequencing. GAPDH as the internal control. Error Bar: Standard Deviation of the mean expression values of Real-time PCR.
Fig 3.
Heatmap showing the Euclidean distances between the samples as calculated from the variance stabilizing transformation of the count data by DEGseq pakage [33].
Nonc1: Non-activated oncosphere 1; Nonc2: Non-activated oncosphere2; Aonc: Activated oncosphere; 4Wmet: 4-week metacestodes miniature vesicles; Cmet: Metacestodes small vesicles cultivated in vitro.
Fig 4.
Analyses of differentially expressed genes (DEGs) among non-activated oncophere (Nonc), activated oncosphere (Aonc) and early stage of metacestode (Emet).
Table 3.
The antigen homologues matched to E. multilocularis proteome.
Fig 5.
Phylogenetic analyses of diagnostic antigen gp50 amino acid sequences.
The neighbor-joining tree was constructed by MEGA 6.0 (www.megasoftware.net); bootstrap values were obtained from 1,000 replicates. Nonc: RPKM>100 at stages of non-activated oncosphere; Aonc: RPKM>100 at stages of activated oncosphere; P_gravid: FPKM>100 at stage of pre-gravid [23]; Gravid: FPKM>100 at stage of gravid [23].
Fig 6.
Protein alignment of putative Em-TSP3 isoforms with four transmembrane.
Fully conserved residues are marked with (*), those replaced with amino acids of strongly similar properties with (:) and of weakly similar properties with (.) LEL variable region are in the solid line box and predicted transmembrane region are in the dashed line box. Protein mutant of the forth transmembrane is shaded in grey. There are three Em-TSP3 isoforms predicted, two of them are conserved with previous study [23], but one of them are intermediate type of the former two isoforms and need further verification.