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Figure 1.

Length distribution of transcripts obtained from O. vulgaris hemocytes transcriptome library.

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Figure 2.

Frequency of contigs showing similarity to known proteins in the NCBI database.

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Table 1.

Summary statistics of sequencing and assembly for O. vulgaris hemocytes transcriptome.

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Figure 3.

Top 35 hit sequences matching O. vulgaris assembled sequences.

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Figure 4.

Distribution of second level GO annotation in three categories: (A) cellular component, (B) molecular function and (C) biological process.

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Figure 5.

Complement and clotting pathway.

Brown figures indicate proteins identified in the O. vulgaris library; and blue figures indicate the absent ones. C1q: Complement C1q binding protein; C1R: C1r subcomponent; C1S: Complement C1 subcomponent; C2: Complement component 2; C4: Complement component 4; C3: Complement component 3; C3a: anaphylatoxin subcomponent 3a; C3b: Opsonin subcomponent 3b; C5: Complement component; C3R: C3 receptor; C5R: C5 receptor; MBL: Manose-binding lectin; MASP1/2: Mannan-binding lectin serine protease 1/2; F12: factor 12; F11: Factor 11; α2M: Alpha-macroglobulin; F2,3,5,7,8,9,10: Coagulation factors 2,3,5,7,8,9,10; MPC: CD46, membrane cofactor protein; VWF: Von Willebrand factor; KLKB1: kallikrein B1; PROC: protein C; PROS1: protein S (alpha); THBD: trombomodulin.

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Figure 6.

TLR/NF-κB signaling pathway.

Brown rectangles indicate proteins identified in the present Octopus vulgaris library and blue rectangles indicate the absent ones. Brown rectangles with red letters indicate proteins in the NF-κB pathway. AKT: RAC-alpha serine/threonine-protein kinase; API1: Transcription factor AP-1; Casp8: Caspase 8; FADD: FAS-associated via death domain; IκB: Inhibitor of NF-κB; IKKε: Inhibitor of nuclear factor kappa-B kinase subunit epsilon; IRAK4: Interleukin-1 receptor-associated kinase 4; IRF3: Interferon regulatory factor 3; IκBα: NF-kappa-B inhibitor alpha; JNK: c-Jun N-terminal kinase; MEKK1: Mitogen-activated protein kinase knase 1; MKK4/6: Mitogen-activated protein kinase kinase 4/6; MyD88: Myeloid differentiation primary response protein 88; Mtor: Mechanistic target of rapamycin; NF-Κb: Nuclear factor kappa-B; PI3K: Phosphatidylinositol 3 kinase; PIM1: Proto-oncogene serine/threonine-protein kinase pim-1; p105: Nuclear factor NF-kappa-B p105 subunit; RAC1: Ras related C3 botulinum toxin substrate; Stat-1: Signal transducer and activator of transcription 1; SOCS-2/5: Suppressor of cytokine signaling; TAB1: TAK1-binding protein1; TAK1: TGF-beta activated protein kinase kinase 1; TIRAP: Toll-interleukin 1 receptor domain-containing adaptor protein; TLR2: Toll-like receptor 2; TLR4: Toll like receptor 4; TOLLIP: Toll interacting protein (├ direct inhibition); TRAF3: TNF receptor-associated factor 3; TRAF6: TNF receptor-associated factor 6; MAVS: Mitochondrial antiviral signaling protein that activates NF-kappa B and IRF 3; INFα/β: Interferon alpha/beta receptor; IRAK1–2: Interleukin receptor associated kinase 1, 2; IRF7: Interferon regulatory factor. P38MAPK: p38 mitogen-activated protein kinases; ECSIT: Evolutionarily conserved signaling intermediate in Toll pathways.

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Figure 7.

Apoptosis pathway.

Green ellipse indicates proteins identified in the present O. vulgaris library and blue ones indicate absence. (├ direct inhibition). AKT/PKB: RAC-alpha serine/thereonine-protein kinse/Protein kinase B; AIF: Apoptosis-inducing factor 1 mitochondiral; ATM: Ataxia telangiectasia mutated protein; BAX: Apoptosis regulator BAX. Bcl2: Apoptosis regulator Bcl-2; Bcl-XL: Bcl-2 like protein 1; BI1: BAX inhibitor-1; Casp 3, 6, 7, 8, 10: Caspase 3, 6, 7, 8, 10; Cytc: Cytochrome C; DFF40, 45: DNA fragmentation factor of 40 kD, 45 kD; FADD: FAS-associated via death domain; IAP: Inhibitor of apoptosis; IKK: Inhibitor of nuclear factor kappa-Bkinase; IκBα: MyD88: Myeloid differentiation primary response protein MyD88; NF-kappa-B inhibitor alpha. IL3R: Interleukin 3 receptor; NFκB: Nuclear factor kappa-B; PI3K: Phosphatidylinositol 3-kinase; p53: Tumor suppressor p53. RIP1: Receptor interacting serine/threonine-protein kinase1; TRADD: TNF receptor superfamily 1 alpha-associated via death domain; TRAF2: TNF-receptor-associated factor 2; TRAIL: TNF-related apoptosis-inducing ligand; Apaf1: Apoptotic protease-activating factor; FLIP: FADD-like apoptosis regulator; PTEN: Phosphatidylinositol-3, 4, 5-trisphosphate 3 phosphatase and dual specificity protein phosphatase PTEN; Smac: Second mitochondria-derived activator of caspase; Chk1/2: Checkpoint kinases 1, 2.

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Figure 8.

Fold change in gene expression analysis by RT-qPCR.

Tissue expression profiles of immune genes in O. vulgaris. Data represent the fold change in expression of the analyzed transcripts relative to β-acting transcript level of sick octopuses (highly infected by A. octopiana), referred to healthy octopuses (null or lowly infected by A. octopiana). Results are mean ± standard deviation. Asterisks denotes significant differences (P<0.05).

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Table 2.

Primer sequences used for RT-qPCR.

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