asap1 morphants exhibit altered susceptibility to Mm infection. (A,B) Western blotting (A) and qRT-PCR (B) analysis of the knockdown efficiency of asap1 morphants in 1, 5, and 10 dpf zebrafish. *P < 0.05, **P < 0.01 vs. control morphants using Student's unpaired t-test. The antibody was against both Asap1a and Asap1b. Data shown are representative of 3 independent experiments. (C) Schematic diagram of the yolk injection method. (D,E) Representative fluorescence images and quantification of bacterial burdens by fluorescence pixel counts of controls, asap1 morphants, or asap1 morphants treated with asap1 mRNA at 5 dpi with equivalent bacterial inocula by the yolk injection method (scale bar 500 μm); *P < 0.05, P-values calculated by Kruskal-Wallis with Dunn's post-test. n (MOs-NC group) = 27, n (MOs-asap1 group) = 27, n (MOs-asap1+asap1mRNA group) = 20. Mean ± SEM from four pooled independent experiments. (F) Schematic diagram of the intravenous injection method. (G,H) Representative fluorescence images and quantification of bacterial burdens by fluorescence pixel counts of controls, asap1 morphants or asap1 morphants treated with asap1 mRNA at 5 dpi with equivalent bacterial inocula by the intravenous injection method (scale bar, 500 μm); **P < 0.01, NS, not significant, P-values calculated by Kruskal-Wallis with Dunn's post-test. n (MOs-NC group) = 18, n (MOs-asap1 group) = 19, n (MOs-asap1+asap1mRNA group) = 20. Each dot represents one larva. Mean ± SEM from three pooled independent experiments. MOs, morpholino oligonucleotide mixture. asap1 means asap1a and asap1b.
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