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Fig. 6 Selenoprotein-N null myoblasts show impairment in metabolism after differentiation at high cell seeding confluency. (A) Immunoblot shows absence of ~ 65 kDa SelN in three selenon-null myoblast lines: C2C12 Exon 3, C2C12 Exon 5, and mouse quadriceps primary cells (Quad PC). α-Tubulin was used as a loading control and is present at ~ 52 kDa in all lines. (B) Results from seahorse cell respirometer show changes in Oxygen Consumption Rate (OCR) and Extra Cellular Acidification Rate (ECAR) parameters at different cell plating densities of selenon-null C2C12 Exon 5 cells. (C) Quantification of OCR and ECAR in increasing concentrations of C2C12 selenon-null exon 5 myoblasts show significant differences when compared to wild type only at 20,000 cell density (N = 10). (D) Quantification of basal OCR and ECAR in C2C12 selenon-null exon 3 and one mouse quadricep primary selenon-null cell lines shows impaired metabolism in KO myoblasts when compared to WT at 20,000 cell density (N = 30). (E) Quantification of basal OCR in three 1 dpf selenon-mutant zebrafish embryos show no differences in metabolism in mutants when compared to WT (N = 48). “**” = p < 0.01, “***” = p < 0.001, “****” = p < 0.0001