Zebrafish selenon mutants show absent or partial expression of Selenoprotein-N. (A) Sanger sequencing chromatograms show analysis of the selenon gene exon 2 for wild type (WT), selenoncl502, selenoncl503, and selenoncl504 homozygotes using genomic DNA from zebrafish tail clips. (B) Real Time qPCR analyses show selenon transcript levels at 1 dpf and 6 dpf in zebrafish knock outs (KO’s) and mutants (Mt): selenoncl502, selenoncl503, and selenoncl504with their correspondent WT. (N = 5 per group) “*” = p < 0.05, “**” = p < 0.01, “***” = p < 0.001, “****” = p < 0.0001. (C) Western Blot analysis shows SelN expression at the predicted size of ~ 65 kDa in positive control (SelN-transfected HEK cells) but not in negative control (WT HEK cells). Protein lysates from 2 dpf zebrafish selenon mutants and their corresponding WT controls show SelN expression in all WT fish, no expression in selenoncl502 and selenoncl503, and reduced expression in selenoncl504 mutant line. (N = 30 per group) (D) Ponceau staining in western blot used to demonstrate equal protein loading throughout the blot
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