Figure 6.

Distribution of transgenic versus endogenous ATP8

Denaturing PAGE western blots of mouse liver tissue from 6-, 12-, 30-, and 50-week-old transgenic C57BL/6J(mtC57BL/6J) mice (A left panel) and C57BL/6J(mtFVB) mice (A, right panel). The immunoblot bands of oATP8 and endogenous ATP8 were quantified by densitometry analysis (using ImageJ), normalized to GAPDH (B) and the ratio of oATP8 and endogenous ATP8 were calculated (C). Denaturing PAGE western blots of mouse liver mitochondria from non-transgenic and transgenic C57BL/6J(mtC57BL/6J) and C57BL/6J(mtFVB) mice (D). The immunoblot bands of oATP8 and endogenous ATP8 were quantified by densitometry analysis (using ImageJ), normalized to aconitase (E) and the ratio of oATP8 and endogenous ATP8 were calculated (F). Approximately fifty micrograms of protein was run per lane (n = 3 animals per group). The animal ID alongside the gender (“M” for male and “F” for female) is listed at the top of the lanes. FLAG-tagged oATP8 protein was immunodetected with mouse anti-FLAG antibody and endogenous ATP8 was immunodetected with rabbit anti-ATP8 antibody. GAPDH and ACO2 were used as loading controls for nuclear and mitochondrial fractions. Error bars show SEM. (B) Two-way ANOVA with Šídák’s multiple comparisons: p > 0.05, ∗∗p = 0.0018, ∗∗∗∗p ≤ 0.0001. (C) Two-way ANOVA with Šídák’s multiple comparisons: p > 0.05, ∗p = 0.0205, ∗∗p = 0.00015. (E) Two-way ANOVA with Šídák’s multiple comparisons: p > 0.05, ∗p = 0.0114. (F) Unpaired t test with Welch’s correction. p > 0.05; NS, not significant.