Flotillin 2

The primary antibodies used in this study included (i) a mouse monoclonal antibody (α5, DHSB, Iowa City, IA, USA) raised against the avian NKA α-subunit (Lin et al., 2003; Yang et al., 2015) , (ii) a mouse monoclonal antibody (flotillin-2, Santa Cruz, CA, USA) raised against the amino acids 150-240 of human flotillin-2 (XP_016879883.1) with approximately 83% similarity with milkfish flotillin-2 (XP_030648347.1) and 80% similarity with tilapia flotillin-2 (XP_003456181.1), and (iii) a rabbit polyclonal antibody (MDBio, Taipei, Taiwan) raised against the specific epitope (AEMPADSGYPAYLGARLA) of the pufferfish (Tetraodon nigroviridis) VHA A (ABX80240.1) with 100 % similarity with milkfish VHA A isoform1 and isoform 2, as well as tilapia VHA A (BAF94024.1). The sequences of two isoforms of VHA A found in the milkfish transcriptome database (Hu et al., 2015) were showed in Fig. S1. The specificities of the VHA A antibody to gills of tilapia and milkfish were tested and showed in Fig. S2. According to the primary antibodies, the secondary antibodies for immunoblots included the horseradish peroxidase-conjugated (i) rabbit anti-mouse IgG, (ii) goat anti-rabbit IgG, and (iii) rabbit anti-goat IgG (GeneTex, Irvine, CA, USA). The similarity between flotillin-2 with milkfish.