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. 2015 Jun 5;5:10903. doi: 10.1038/srep10903

Figure 4. Increased expression of autophagic markers in Gaucher fibroblasts.

Figure 4

(a) Quantification of acidic vacuoles in control and Gaucher fibroblasts by LysoTracker staining and flow cytometry analysis. (b) Representative LysoTracker staining images in control and Gaucher fibroblasts. Scale bar= 15 μm. (c) Effect of CoQ, PC and CoQ + PC on the amount of acidic vesicules in Gaucher-B fibroblasts. Control and Gaucher fibroblasts were cultured in the presence or absence of CoQ (25 μM), PC NAdBT-AIJ (25 μM) or CoQ + PC (25 μM + 25 μM) for 96 h. Acidic vacuoles were quantified by LysoTracker staining and flow cytometry analysis. For control cells, the data are the mean ± SD for experiments conducted on 2 different control cell lines. Data represent the mean ± SD of 3 separate experiments. *p < 0.01 between control and Gaucher fibroblasts. ap < 0.05 between the presence and the absence of CoQ. bp < 0.05 between the presence and the absence of PC. cp < 0.05 between the presence and the absence of CoQ + PC. #p < 0.05 between CoQ + PC and CoQ or PC treatment. (d) The expression levels of LC3-I (upper band) and LC3-II (lower band), ATG12, BECLIN1 and cathepsin D were determined in the control and Gaucher-B fibroblast cultures by Western blotting. The ATG12 band represents the Atg12-Atg5 conjugated form. Fibroblast cultures were grown in normal culture medium or in medium supplemented with CoQ (25 μM), PC (25 μM) or the combination of both CoQ + PC (25 μM + 25 μM) for 96 h. Actin was used as a loading control. The densitometric analysis of Western blottings is showed in Supplementary Figure S6.