Supplementary figures,table, and captions

Fig. S1

Caption Fig. S1:

Total RNA was extracted from gills, stomach, intestine, skin, kidney, head kidney, spleen, muscle, heart, brain, blood, and liver of healthy cobia, respectively. RNA sample concentration was assessed with a NanoDrop spectrophotometer prior to the reverse transcription (RT). The first-strand cDNA was synthesized from 2 μg of DNase-treated total RNA. The same amount cDNA was used as template for PCR reactions. RT-PCR was used to detect the expression levels of β-actin and RcCC2 mRNA in various tissues of non-stimulated cobia. M, DL2000 DNA marker; Gi, gills; St, stomach; In, intestine; Sk, skin; Ki, kidney; Hk, head kidney; Sp, spleen; Mu, muscle; He, heart; Br, brain; Bl, blood; Li, liver.

Fig. S2

（A）

（B）

（C）

Caption Fig. S2:

Total RNA was extracted from liver of cobia prior to injection (0h) and at 3, 6, 12, 24, 48, and 72 h post-injection.RNA sample concentration was assessed with a NanoDrop spectrophotometer prior to the reverse transcription (RT). The first-strand cDNA was synthesized from 2 μg of DNase-treated total RNA. The same amount cDNA was used as template for PCR reactions. RT-PCR was used to detect the expression levels of β-actin and RcCC2 mRNA in liver at various times after injection with PBS (A), poly(I:C) (B) or inactivated bacterial vaccine (C).

Fig. S3

Caption Fig. S3:

The cDNA and deduced amino acid sequences of RcCC2 from cobia. The nucleotide sequences are on the upper lines and putative amino acid sequences are in the lower lines. The start and stop codon are boxed. The four conserved cysteines (C) are marked by triangles. The predicted cleavage of the signal sequence to the putative mature protein, two cytokine RNA instability motifs (ATTTA) , and a polyadenylation signal (AATAAA) are underlined.

Table S1

Accession numbers / Species / Chemokine
name / Amino acid
Identities (%) / Accession numbers / Species / Chemokine
name / Amino acid identities (%)
P22362 / Human / CCL1 / 18.0 / DQ173276 / Catfish / SCYA101 / 15.3
P13500 / Human / CCL2 / 15.3 / DQ173280 / Catfish / SCYA106 / 34.2
P10147 / Human / CCL3 / 14.4 / DQ173286 / Catfish / SCYA112 / 20.7
P13236 / Human / CCL4 / 15.3 / DQ173287 / Catfish / SCYA113 / 21.6
P13501 / Human / CCL5 / 15.3 / DQ173289 / Catfish / SCYA115 / 15.3
P80098 / Human / CCL7 / 15.3 / DQ173290 / Catfish / SCYA116 / 15.3
P80075 / Human / CCL8 / 15.3 / DQ173293 / Catfish / SCYA119 / 22.5
P51671 / Human / CCL11 / 11.7 / DQ173295 / Catfish / SCYA121 / 9.0
Q99616 / Human / CCL13 / 14.4 / DQ173298 / Catfish / SCYA126 / 16.2
Q16627 / Human / CCL14 / 14.4 / AAF23867 / Rainbow trout / CK1 / 19.8
Q16663 / Human / CCL15 / 16.2 / AAM09300 / Rainbow trout / CK2 / 13.5
O15467 / Human / CCL16 / 14.4 / AJ315149 / Rainbow trout / CK3 / 15.3
Q92583 / Human / CCL17 / 18.0 / CA371157 / Rainbow trout / CK4A / 18.9
P55774 / Human / CCL18 / 14.4 / CA383670 / Rainbow trout / CK5A / 14.4
Q99731 / Human / CCL19 / 22.5 / CA378686 / Rainbow trout / CK9 / 11.7
P78556 / Human / CCL20 / 18.9 / CA361535 / Rainbow trout / CK10 / 39.6
O00585 / Human / CCL21 / 24.3 / CA346383 / Rainbow trout / CK12B / 23.4
O00626 / Human / CCL22 / 16.2 / ADE58983 / Gilthead seabream / CK1 / 18.9
P55773 / Human / CCL23 / 15.3 / ADE58986 / Gilthead seabream / CK3 / 17.1
O00175 / Human / CCL24 / 17.1 / ADE58984 / Gilthead seabream / CK5 / 14.4
O15444 / Human / CCL25 / 10.8 / ADE58987 / Gilthead seabream / CK7 / 16.2
Q9Y258 / Human / CCL26 / 14.4 / ADE58985 / Gilthead seabream / CK8 / 18.9
Q9Y4X3 / Human / CCL27 / 14.4 / ADE58982 / Gilthead seabream / CK10 / 57.7
Q9NRJ3 / Human / CCL28 / 18.0 / JF975591 / Cobia / RcCC1 / 17.1
P27784 / Mouse / CCL6 / 9.9 / JF975592 / Cobia / RcCC3 / 18.0
P51670 / Mouse / CCL9 / 15.3 / ABC69047 / Turbot / KC70 / 56.8
Q62401 / Mouse / CCL12 / 16.2 / AEL31667 / Half-smooth tongue sole / CsCCK1 / 43.2

Caption Table S1:

Amino acid identity of RcCC2 with mammals and other fish CC chemokines.