Fig. 6.

Protein expression of the mitophagic marker, Fundc1, in 4 and 10 week WT and KO littermates, and assessment of autophagic/mitophagic flux upon Bafilomycin A1 (BAF) treatment in the LV tissue of 10 week frataxin KO mice versus WT littermates. (A, B) Representative immunoblots and densitometry showing Fundc1 expression in (A) total cellular protein, and (B) mitochondrial fractions, of 4 and 10 week KO versus WT littermates. Gapdh was used as a loading control for total protein-loading in (A), while Hsp90 and Tomm20 were used as protein-loading controls for cytoplasmic and mitochondrial fractions in (B), respectively. In (A, B) data are mean ± SEM (n = 4–6). Unpaired two-tailed Student's t-tests were used. (C) BAF inhibits autolysosomal degradation of autophagic components, i.e., Lc3-II and p62. Representative (i) immunoblots and (ii-vi) densitometry of (ii) Lc3-I, (iii) Lc3-II, (iv) Lc3-II:Lc3-I ratio, (v) p62, and (vi) Mfn1, in the heart from 10 week WT and KO littermates after treatment for 30 min with or without BAF (6 mg/kg; i.p.). Data are mean ± SEM (n = 6 mice/genotype). One-way ANOVA with Tukey's post hoc tests were used. *, p < 0.05; **, p < 0.01; ***, p < 0.001 versus WT. #, p < 0.05; ##, p < 0.01 versus vehicle control. a.u., arbitrary unit. n.s., not significant.