한수지 50(3), , 2017 Original Article Korean J Fish Aquat Sci 50(3), ,2017 조피볼락 (Sebastes schlegelii) Interferon Regulatory Factor 8 (IRF8)

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한수지 50(3), 302-310, 2017 Original Article Korean J Fish Aquat Sci 50(3),302-310,2017 조피볼락 (Sebastes schlegelii) Interferon Regulatory Factor 8 (IRF8) 의분자유전학적특성및발현분석 양혜림 권혁재 이성도 S.D.N.K Bathige 김명진 이제희 제주대학교해양생명과학과 Molecular Characterization and Expression Analysis of Interferon Regulatory Factor 8 (IRF8) in the Black Rockfish Sebastes schlegelii Hyerim Yang, Hyukjae Kwon, Seongdo Lee, S.D.N.K Bathige, Myoung-Jin Kim, and Jehee Lee Department of Marine Life Sciences, Jeju National University, Jeju 63243, Korea Interferon regulatory factor 8 (IRF8) is essential for the development of B and T cells, as well as for the activity of dendritic cells and macrophages. We performed molecular characterization of IRF8 from rock fish, Sebastes schlegelii (Ss), and investigated the spatial and temporal profile of mrna expression after challenge with lipopolysaccharide (LPS), polyinosinic:polycytidylic acid (poly I:C), or Streptococcus iniae. The full-length cdna sequence of SsIRF8 was 1,657 bp, containing an ORF of 1,266 bp. The gene had a predicted molecular mass of 47.7 kda and an isoelectric point of 5.99. The amino acid sequence coded by this gene showed the highest degree of identity (90.8%) and similarity (96.2%) with IRF8 from Oplegnathus fasciatus. The SsIRF8 mrna was expressed ubiquitously, at varying levels, with the highest level of expression observed in the spleen. To confirm the role of SsIRF8 in mediating the immune response, we measured SsIRF8 mrna expression in the splenic tissue at different time points after injection with LPS, poly I:C, or S. iniae. The qrt-pcr results showed that SsIRF8 mrna expression in the poly I:C-injected group was highly upregulated 6 hr after exposure (P<0.05). Expression of SsIRF8 mrna in the S. iniae-injected group peaked at 24 hr. These results suggest that SsIRF8 might be important in regulating the strength of the rockfish immune response to immunostimulatory agents. Key words: Black rockfish, IRF8, Immune regulator, mrna expression 서론 Interferon regulatory factor (IRF) family,,, (Barnes et al., 2002 ), 11 IRF family (Huang et al., 2010). IRF (interferon, IFN) interferon stimulated genes (ISGs) (Nguyen et al., 1997),,,, (Pfeffer et al., 1998). Type I Type II IFN (Inkpen et al., 2015), Type I IFNs (IFN IFN ) Type I IFNs (Mamane et al., 1999; Taniguchi et al., 2001; Tailor et al., 2007). IRF family N- ~120 amino acid 5 tryptophan (Trp) helixturn-helix motif DNA binding domain (DBD) (Nguyen et al., 1997). IRF DBD consensus DNA sequence IFN-stimulated response element (ISRE) GAAA, AANNNGAA ISRE (Honda and Taniguchi, 2006; Paun and Pitha, 2007). IRF-associated domain (IAD) IRF family C- IRF3-10 IAD1, IRF1 IRF2 IAD2 (Mamane https://doi.org/10.5657/kfas.2017.0302 Korean J Fish Aquat Sci 50(3) 302-310, June 2017 This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial Licens (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. Received 1 June 2017; Revised 22 June 2017; Accepted 23 June 2017 Corresponding author: Tel: +82. 64. 754. 3473 Fax: +82. 64. 756. 3493 E-mail address: jehee@jejunu.ac.kr (J. Lee), mj.kim.lucky@gmail.com (M-J. Kim) Copyright 2017 The Korean Society of Fisheries and Aquatic Science 302 pissn:0374-8111, eissn:2287-8815

IRF8 의특징및발현분석 303 et al., 1999). IAD IRFs homodimer heterodimer (Eroshkin and Mushegian, 1999; Ozato et al., 2007). IRF family N- DBD C- IAD DBD IAD (Nehyba et al., 2002). IAD IRF family, (IRF1, IRF3, IRF7, IRF9), (IRF2, IRF8), (IRF2, IRF4, IRF5, IRF8) (Stellacci et al., 2004). IRF1, IRF2, IRF8 myeloid lymphoid cell (Wang and Morse, 2009; Marecki et al., 2015)., IRF8 IRF4 hematopoietic cell, myeloid progenitor cell B T (Lu, 2008), dendritic cells (DCs) plasmacytoid dendritic cells (pdcs) type I IFNs (Alter-Koltunoff et al., 2008). IRF8 Ets-IRFE motif ISRE motif (Lee et al., 2006). IRF8 LPS-TLR4 polyinosinic:polycytidylic acid (poly I:C)-TLR3 Toll-like receptor IFN- signaling pathways (Zhao et al., 2006), poly I:C-activated signaling pathways plasmacytoid DCs (pdcs) IFN DCs subtype IRF8 (Gabriele and Ozato, 2007; Tailor et al., 2007). IRF8 rainbow trout (Holland et al., 2010), turbot (Chen et al., 2012), rock bream (Bathige et al., 2012), Japanese flounder (Hu et al., 2013), tongue sole (Zhang et al., 2015), atlantic cod (Inkpen et al., 2015). in silico, IRF8, LPS, poly I:C, Streptococcus iniae. 재료및방법 Rockfish 의 IRF8 cdna sequence 분석 454 GS FLX sequencing (Roche 454 Life science, USA) cdna sequence database. DNAsist 2.2 software open reading frame (ORF). Expert Protein Analysis System ProtParam tool (SIB Swiss Institute of Bioinformatics, Swiss) (http://web. expasy.org/protparam)., Basic Local Alignment Search Tool (BLAST) algorithm SsIRF8 IRF family member. IRF family domain motif proteins, motif scan Pfam hidden Markov models (Local models) (SIB Swiss Institute of Bioinformatics, Swiss) (http://hits.isb-sib.ch/cgi-bin/ PFSCAN) IRF8 IRF8 homolog Matgat software ClustalW2 server (EMBL- EBI, UK) (http://www.ebi.ac.uk/tools/clustalw2) pairwise sequence alignment multiple sequence alignment strategies. Molecular Evolutionary Genetic Analysis (MEGA) software version 7.0 neighbor-joining (NJ) 1,000 bootstrap. 시험어준비및조직분리 (200 20g) 22 1 400 L. SsIRF8. gill, heart, head kidney, spleen, liver, intestine, kidney, testis, ovary 9-80., 0.2% heparin sodium salt (USB, USA) (-1 ml/fish). (4, 3,000 g, 10 ) peripheral blood cells (PBCs) -80. 공격실험을통한면역자극비교및조직분리, SsIRF8 200 L 1 phosphate buffered saline (PBS) lipopolysaccharide (LPS; 1.25 g/ L) 200 L 1 PBS polyinosinic: polycytidylic acid (poly I:C; 1.5 g/ L, Sigma, USA), 200 L 1 PBS Streptococcus iniae (1 10 5 colony-forming units/ L). 200 L 1 PBS 3, 6, 12, 24, 48, 72 5 RNA -80.

304 양혜림ㆍ권혁재ㆍ이성도ㆍ S.D.N.K Bathige ㆍ김명진ㆍ이제희 RNA 추출및 cdna 합성 qpcr 에의한발현분석 QIAzol (Qiagen) RNA. RNA purity 1.5% agarose gel, 260 nm Drop Plate (Thermo Scientific, Waltham, MA, USA). cdna RNA 2.5 g reaction mixture 20 L PrimeScript first-strand cdna synthesis kit (TaKaRa, Shiga, Japan)., cdna nuclease-free water 40 qpcr assay 20. quantitative real-time PCR (qpcr) IRF8 mrna. Reference gene elongation factor 1- (EF1 ) forward (5'-AACCTGACCACTGAGGTGAAGTCTG-3'), Reverse (5'-TCCTTGACGGACACGTTCTTGATGTT-3') oligonucleotide primer. SsIRF8 oligonucleotide primer forward (5'-CATCAATGCTTTCCCTCC- GCATCAAG-3'), reverse (5'-TGTGCATCAGCTTTCCTCC- GTAGT-3'). qpcr Real Time System TP800 Thermal Cycler Dice (TaKaRa). 5 -UTR(67) AAGAGCTCAACCGCTTATCACACTGCACAAGGAAACAATCCCCAAAACGCCCTTTCTCTTCATTAAG ATG TCA AAC ACT GGA GGT CGG AGA CTG AAG CAG TGG CTG GTG GAG CAG ATC CAG AGT GGC CAG TAC TCT GGA M S N T G G R R L K Q W L V E Q I Q S G Q Y S G CTG CAG TGG GAG GAT GAC AGC CGC ACT ATG TTC AGA ATC CCA TGG AAA CAC GCA GGC AAA CAA GAC TAC AAC L Q W E D D S R T M F R I P W K H A G K Q D Y N CAA GAA GTA GAC GCA TCC ATT TTC AAG GCC TGG GCT GTG TTT AAA GGC AAG TTT AAG GAG GGG GAG AAG GCT Q E V D A S I F K A W A V F K G K F K E G E K A GAG CCT GCC ACC TGG AAG ACC AGA CTC CGC TGT GCC CTG AAT AAA AGC CCT GAC TTT GAG GAG GTG ACT GAA E P A T W K T R L R C A L N K S P D F E E V T E AGG TCG CAG CTG GAC ATC TCT GAG CCC TAT AAA GTC TAC CGC ATT GTA CCT GAG GAA GAG CAG AAA AAC GGC R S Q L D I S E P Y K V Y R I V P E E E Q K N G AAA GGC GCA TTG ATG GCC ATG GCA GCC ACC ACC AGC TCC GGT GAT ATC AAT GAC ATG GAC TGC AGC CCT GCA K G A L M A M A A T T S S G D I N D M D C S P A GAG ATA GAG GAG CTC ATC AAA GAG GAG GAA GGC TGT AGT ATC CAG ACC AGT CCA GAG TAT TGG TCC CAT GGC E I E E L I K E E E G C S I Q T S P E Y W S H G AGC ATC AAT GCT TTC CCT CCG CAT CAA GAC CCT TTG CCA TCA GGC AAT CTC AGC ACA GCT TTC TCC CAG ATG S I N A F P P H Q D P L P S G N L S T A F S Q M ATG ATC AGT TTC TAC TAC GGA GGA AAG CTG ATG CAC AAC ACA CAT GTT GTT CAT CCT GAA GGC TGC CGA ATC M I S F Y Y G G K L M H N T H V V H P E G C R I GCC CCA CAA CAG CAC CTG GGC CGT GGC GCC CTA TAC AAT TCA GAC AGC ATG CAG AGT GTT CAC TTT CCT CCC A P Q Q H L G R G A L Y N S D S M Q S V H F P P GCT GAG CAC ATC GAG TAC GAC CGC CAG CGC CAT GTC ACA CGC AAG CTC CTG GGA CAC CTG GAG AGA GGT GTA A E H I E Y D R Q R H V T R K L L G H L E R G V CTG GTC CGT GCC AAC CAA GAG GGC ATC TTC ATC AAA AGG CTG TGC CAG AGC CGT GTC TTC TGG AGC GGG TTG L V R A N Q E G I F I K R L C Q S R V F W S G L GGA GAA GTG GGC TCA CAA TAT GGC TCC ATG CCT TGT AAA CTT GAG AGG GAT GCT GTA GTC AAG ATT TTT GAC G E V G S Q Y G S M P C K L E R D A V V K I F D ACA GGA AAG TTT CTT CAA GCT CTC CAG CTG TAC CAG GAG GGT CAG TTT CCA GCT CCT GAT CCG ACA GTG ACT T G K F L Q A L Q L Y Q E G Q F P A P D P T V T CTG TGT TTC GGA GAG GAG CTC CAT GAC CTC AGC AAT GCC AAG AGC AAG CTG ATC ATT GTG CAG ATC ACT GTG L C F G E E L H D L S N A K S K L I I V Q I T V GTG AAC TGC CAG CAG CTG CTG GAG GCA GTG AAC ATG CGT CGC TCC CAG CCT TAC TCC AAC AGC TCA AAC CTG V N C Q Q L L E A V N M R R S Q P Y S N S S N L GAG ATG TCT GAT AAT GTG GCC ATT GAC CAG ATG GCC CGC ATC TAC CAG GAC TTG TGC AGC TAT AGC GGC CCC E M S D N V A I D Q M A R I Y Q D L C S Y S G P TAC CAG GAC TTG TGC AGC TAT AGC GGC CCC CAG AGG CCA GCC TGC TAC AGG GAC AAC ATG CCC ATC ACT GCC Y Q D L C S Y S G P Q R P A C Y R D N M P I T A 3 -UTR(324) TGAGCCGTGAGCAGCAGCAAAACACCTGCCAGGAGCTCTTCAGCGAGTACCAGACTGACTGTACAACATATTAGGGACAGTCCTTATTTT CCTGACATACACTGACAAGGTGAATTCAGGTAAAAACCAGTATCACAATTAGTTTTGACTTTTTAAATGGAGCAAATATCTTACAGTAAGACT TTTACGCTGGAGCTCAGTATTAGGATTTTGTATACCAGTAATGTTTATGTGTCTTATATGTACAATGCTAACACAAACTCTGTGTCTAACTTC TGCTTTTTTACAATAAGAGAGTGTTATCATCCAGCTCGAGAGAAAAAG Fig. 1. The cdna and amino acid sequences of the black rockfish Sebastes schlegelii IRF8. The open reading frame from ATG to TGA is shown in the middle box. The amino acid sequence of the DNA binding domain (DBD) is shaded in gray and the IRF-association domain 1 (IAD1) is in bold, shaded in gray, and boxed. The five conserved tryptophan (W) residues in the DNA binding domain are boxed.

IRF8 의특징및발현분석 305 qpcr 3 L cdna, 5 L 2 TaKaRa Ex Taq SYBR premix, 0.4 L 10 M forward, reverse primer, 1.2 L H 2 O 10 L. qpcr 95 10 1 cycle, 95 5, 58 10, 72 20 35 cycle, 95 15, 60 30, 95 15 1 cycle 3. 통계학적분석 SPSS version 18 program (IBM, USA) one-way ANOVA Table 1. The percentages of identity and similarity of SsIRF8 amino acid sequence with that of other species. Oplegnathus fasciatus IRF8 protein sequence showed the highest values Scientific Name Accession Number Sebastes schlegelii Oplegnathus fasciatus Epinephelus coioides Epinephelus coioides Miichthys miiuy Dicentrarchus labrax Scophthalmus maximus Paralichthys olivaceus Larimichthys crocea Gadus morhua Oncorhynchus mykiss Ctenopharyngodon idella Gymnocypris przewalskii Danio rerio Scleropages formosus Gallus gallus Cathartes aura Sebastes schlegelii Oplegnathus fasciatus Epinephelus coioides Epinephelus coioides Similarity (%) Identity (%) 90.8 90.5 90 90.5 90 89.1 85.8 87 73.4 73.3 67.3 64.8 64.5 64.5 51.9 51.8 AFU81290.1 96.2 91.5 91 92.9 92.7 90.3 87 89.3 71.1 72.4 67.7 65.7 65.9 64.4 52.6 52.3 AND67453.1 95.5 96.9 99.1 91.2 91 89.3 86.5 87.7 72 71.5 66.4 64.3 65.2 63.7 51.9 52.3 AND67452.1 95 96.4 99.5 90.8 90.5 88.9 86 87.2 71.8 71.3 66.1 64.5 64.8 63.4 51.9 52.3 Miichthys miiuy AHB59740.1 95.3 96.4 96 95.5 94.1 88.6 85.3 95 72.7 72.4 67.5 65.7 65.5 64.6 52 52.5 Dicentrarchus labrax Scophthalmus maximus Paralichthys olivaceus Larimichthys crocea AKC57349.1 95.7 97.6 96.9 96.4 97.9 88.9 85.8 90.5 73.1 73.1 67.7 66.1 64.8 64.4 52.1 52.1 AFE88896.1 94.3 95.7 95 94.5 95 96.4 88.8 85.8 71.3 71 66.2 64.8 64.6 62.2 52.5 51.6 AFE18694.2 93.6 94.3 94.8 94.3 93.6 94.1 95 82.5 69.4 70.2 64.4 63.5 61.9 63 52.2 50.7 KKF19617.1 91.5 92.9 92.7 92.2 95.7 94.3 92.4 91 74.2 70.6 66.6 64.8 63.5 64.3 51.1 51.6 Gadus morhua AJR33029.1 86.5 86.7 86.7 86.3 87.7 87.7 86.5 85.5 87.4 71.6 65.5 64.7 64.5 60.6 51.3 52.8 Oncorhynchus mykiss Ctenopharyngodon idella Gymnocypris przewalskii ALS92677.1 81.8 82 81.8 81.3 82.7 83.8 81.3 80.9 80.6 81.5 71.5 69.9 66 64.6 54.7 55.6 AMT92197.1 80.4 81.5 82.2 81.8 81.8 81.5 79.4 79.4 79 79.9 81.8 92.5 85.1 66 56.5 57.7 AMB19593.1 79.4 80.8 80.8 80.6 80.4 80.4 79.7 78.5 78 79.7 81.8 94.9 82.7 63.2 56.5 57.8 Danio rerio NP_001002622.1 80.4 80.9 80.9 80.4 81.8 82 80.9 79.2 78.7 80.1 78.4 91.1 90 61.6 55.2 56.1 Scleropages formosus KPP68827.1 78.4 77.7 77.7 77.7 78.6 79.1 77.2 77.9 77 77.2 79.1 78.2 77.2 74.7 51.4 52.9 Gallus gallus NP_990747.1 68.9 70.4 70.1 69.9 68.9 69.4 70.1 69.4 68.5 68.9 68.7 72.9 72.2 73.2 68.5 91.1 Cathartes aura KFP53072.1 68 68.9 69.2 68.9 68.7 68.5 67.8 67.8 68.2 68.2 69.6 72.4 72.4 72.2 69.7 96

306 양혜림ㆍ권혁재ㆍ이성도ㆍ S.D.N.K Bathige ㆍ김명진ㆍ이제희, mrna (Mean) (Standard deviation) IRF8 mrna. P<0.05. 결과및고찰 IRF8 full-length cdna sequence mrna. SsIRF8 full-length cdna sequence 1,658 bp, 5 -untranslated region (UTR) 67 bp, 3 -untranslated region (UTR) 324 bp. Open reading frame (ORF) 1,266 bp 422 (Fig. 1). 47.77 kda 5.99. SsIRF8 bioinformatics tool in silico IRF family DBD, tryptophan, IAD. DBD IRF family ISRE/IRF-E consensus sequence. 5 tryptophan DBD IRF motif (Eroshkin and Mushegian, 1999; Nehyba et al., 2002). SsIRF8 DNA-binding domain N- 6-113 aa IRF3-10 IRF-association domain (IAD) C- 189-371 aa. 5 tryptophan 12W, 27W, 39W, 59W, 77W (Fig. 1). IRF8 IRF8, Oplegnathus fasciatus IRF8 96.2%, 90.8% Epinephelus coioides, Miichthys miiuy, Dicentrarchus labrax 95.5%, 95.3%, 95.7% (Table 1). Phylogenetic tree SsIRF8. (Miichthys miiuy), (Larimichthys crocea), (Dicentrarchus labrax), (Oplegna- 65 99 100 80 99 91 69 47 100 31 86 100 Miichthys miiuy Larimichthys crocea Dicentrarchus labrax Oplegnathus fasciatus 100 Epinephelus coioides Epinephelus coioides(2) Scophthalmus maximus Paralichthys olivaceus Sebastes schlegelii Gadus morhua Oncorhynchus mykiss Scleropages formosus Danio rerio Ctenopharyngodon idella Gymnocypris przewalskii Gallus gallus 100 Cathartes aura 0.05 Fig. 2. Neighbor-joining phylogenetic tree of IRF8 constructed with Mega7. The bootstrap confidence values shown at the nodes of the tree are based on 1000 bootstrap replications. Black rock fish Sebastes schlegelii IRF8 is indicated with a black dot ( ) and the scientific name of the fish is in bold. GenBank accession numbers are as follows: Miichthys miiuy, AHB59740.1; Larimichthys crocea, KKF19617.1; Dicentrarchus labrax, AKC57349.1; Oplegnathus fasciatus, AFU81290.1; Epinephelus coioides, AND67453.1, AND67452.1; Scophthalmus maximus, AFE88896.1, Paralichthys olivaceus AFE18694.2; Gadus morhua, AJR33029.1; Oncorhynchus mykiss, ALS92677.1; Scleropages formosus, KPP68827.1; Danio rerio, NP_001002622.1; Ctenopharyngodon idella, AMT92197.1; Gymnocypris przewalskii, AMB19593.1; Gallus gallus, NP_990747.1; Cathartes aura, KFP53072.1.

IRF8 의특징및발현분석 307 Fig. 3. Multiple sequence alignment of SsIRF8 amino acid sequence with other known orthologs from GenBank. The alignment was conducted using ClustalW. The DNA binding domain (DBD) and IRF-association domain1 (IAD1) are shown with arrows. The conserved tryptophan (W) residues in the DBD are boxed and shaded in white. thus fasciatus), (Epinephelus coioides), (Scophthalmus maximus), (Paralichthys olivaceus), (Gadus morhua), (Oncorhynchus mykiss), (Scleropages formosus), (Danio rerio), (Ctenopharyngodon idella), Gymnocypris przewalskii, (Gallus gallus), (Cathartes aura) IRF8, SsIRF8 cluster (Fig. 2). Multiple alignment IRF8 DBD IAD1 homology

308 양혜림ㆍ권혁재ㆍ이성도ㆍ S.D.N.K Bathige ㆍ김명진ㆍ이제희 Relative mrna expression level of SsIRF8 350 300 250 200 150 100 50 0 SP SK IT GL BL HK KD HT LV St MS Tissue type Relative mrna expression of SsIRF8 4.5 4 3.5 3 2.5 2 1.5 1 0.5 0 Poly I:C S.iniae LPS control 3hr 6hr 12hr 24hr 48hr 72hr Fig. 4. SsIRF8 mrna expression in different tissues (SP- spleen, SK-skin, IT-intestine, GL-gill, BL-blood, HK-head kidney, LVliver, St-stomach, MS-muscle) of the healthy black rockfish Sebastes schlegelii as measured using real time qpcr. The relative expressions of the IRF8 are shown as a ratio to IRF8 expression in the MS. Sebastes schlegelii EF1α was used as a reference gene. Data are shown as the mean (n=3) and the error bars represent the standard deviation.. SsIRF8 DBD Oplegnathus fasciatus, Gadus morhua, Oncorhynchus mykiss, Gallus gallus, Mus musculus, Homo sapiens IAD1 (Fig. 3). IRF8 lymphomyeloid-rich (Lehtonen et al., 2003; Wang and Morse, 2009). lymphomyeloid-rich,, IRF8,.,, (Holland et al., 2010; Bathige et al., 2012; Chen et al., 2012; Hu et al., 2013). SsIRF8 9 (gill, heart, head kidney, spleen, liver, intestine, kidney, testis, ovary) PBCs cdna qpcr. Elongation factor 1- (EF1 ) reference gene. qpcr mrna. 256,,,,,, (Fig. 4). myeloid progenitor cell B, T IRF8 (Lu, 2008) SsIRF8, lymphomyeloid-rich, Relative IRF8 expression post Poly I:C, S.iniae, LPS challenge Fig. 5 Gene expression of SsIRF8 in the spleen after challenge with either poly I:C or S. iniae or LPS. All data were normalized to that of Sebastes schlegelii EF1α. The time course of IRF8 mrna expression in the spleen at 0, 3, 6, 12, 24, 48, 72 hr post injection is shown. Data are presented as the mean (n=3) and the error bars represent the standard deviation. Asterisks () represent significant differences (P<0.05).,,, (Hu et al., 2013). mrna PBS, S. iniae, poly I:C, LPS qrt-pcr. Poly I:C 6 0 6 (P<0.05). S. iniae 12 12, 24, 48 24 (P<0.05). LPS 6 12 (P<0.05). SsIRF8 (Fig. 5)., SsIRF8 LPS- TLR4 polyinosinic:polycytidylic acid (poly I:C)-TLR3 Toll-like receptor IFN- signaling pathways IRF8 (Zhao et al., 2006). poly I:C,. poly I:C pathogen associated molecular pattern (PAMP) (Perdiguero and Esteban, 2009; Hu et al., 2013), poly I:C-activated signaling pathways SsIRF8 (Gabriele and Ozato, 2007; Tailor et al., 2007). SsIRF8

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