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Figure 3


Fig. 3. A phylogenetic tree of full-length amino acid sequences of fish and human cation–chloride cotransporters, constructed by the neighbor-joining method. Tilapia NKCC1a, NKCC1b, NKCC2 and NCC are highlighted. Human K+/Cl– cotransporter (KCC1) was used as an outgroup to root the tree. The sequences were grouped into four clades (NKCC1 clade, NKCC2 clade, conventional NCC clade and fish-specific NCC clade). Numbers at the nodes are the bootstrap values for 1000 replications, shown as percentages. Bar, evolutionary distance of 0.05 amino acid substitution per site. The GenBank accession numbers are as follows: tilapia (NKCC1a, AY513737; NKCC1b, AY513738; NKCC2, AY513739; NCC, EU518934), eel (NKCC1a, AJ486858; NKCC1b, AJ486859; NKCC2{alpha}, AJ564602; NKCC2β, AJ564603; NCC{alpha}, AJ564604; NCCβ, AJ564606), zebrafish (zNCCg, EF591989; NCCa, NP_001038545; NCCb, XP_001342888; NCCc, XP_699464), medaka (NCCa, Ensembl protein report for ENSORLP00000000509; NCCb, ENSORLP00000023616; NCCc, scaffold1460 of golw_scaffold Hd-rR 200506), flounder (NCC, L11615), shark (NKCC1, U05958; NKCC2, AF521915) and human (NKCC1, U30246; NKCC2, NM_000338; NCC, X91220; KCC1, NM_005072).