Figure 9.

Effect of riboflavin and CoQ on autophagic markers in MELAS cybrids. (A) Quantification of acidic vacuoles in Wt and MELAS cybrids by LysoTracker staining and flow cytometry analysis. Cells were cultured in the presence or absence of riboflavin (Ribo) or CoQ. Results are expressed in arbitrary units (a.u). (B) Protein expression levels of LC3-I (upper band) and LC3-II (lower band), ATG12 and complex I (30 kDa subunit) were determined in Wt and MELAS (M) cybrid cultures by Western blotting. The ATG12 band represents the Atg12–Atg5 conjugated form. Cybrid cultures were grown in normal culture medium or in medium supplemented with 0.06 µM riboflavin (Ribo) or 100 µM CoQ for 72 h. Actin was used as a loading control. (C) Densitometry was performed using ImageJ software. Actin was used as loading control. Data in arbitrary units (a.u.) represent the mean ± SD of three separate experiments. *P < 0.01 between Wt and MELAS cybrids. ^aP < 0.01, between the presence and the absence of riboflavin or CoQ.