Skip to main content
. 2019 Aug 15;9:11878. doi: 10.1038/s41598-019-48047-x

Figure 5.

Figure 5

The autophagic flux is impaired in sacsin KO cells. (A) Western blotting analysis of LC3 and p62 autophagy markers in WT cells treated for 2 h with the mitochondrial uncoupler FCCP (20 µM) in the absence or the presence of the lysosomal inhibitor bafilomycin A1 (200 nM). The densitometry ratios of LC3-II/LC3-I and of p62 normalized versus GAPDH are reported, and they show correct autophagic flux induction by carbonyl cyanide 4-trifluoromethoxyphenylhydrazone (FCCP) treatment. (B) Western blotting analysis showing LC3 and p62 levels in sacsin KO cells treated for 2 h with the uncoupler FCCP (20 µM) in the absence or the presence of bafilomycin A1 (200 nM). The densitometry ratios of LC3-II/LC3-I and p62 normalized vs GAPDH showed the failure of LC3-II to increase in the presence of bafilomycin, even after normal medium recovery, and p62 accumulation, these findings together indicate a defective autophagy process in KO cells.Data shown in this figure were reproduced independently three times. ns, not statistically significant; *p < 0.05; **p < 0.01; and ***p < 0.001. (Baf = bafilomycin A1; Re = normal medium recovery). Full-length blots are presented in the Supplementary Information 3.