Fig. 8. A77 1726 induces SOD1^G93A co-localization with autophagosomes.

RFP-LC3 stably transfected NSC34 cells were transiently transfected with SOD1-GFP (a) or SOD1^G93A-GFP (b) expression vectors. After incubation for 40 h, the cells were treated with DMSO (0.2%), A77 1726 (200 μM) or rapamycin (50 nM) (a) for 24 h. The cells were fixed and examined under a confocal microscope for the localization of autophagosomes (RFP-LC3) and for SOD1-GFP or SOD1^G93A-GFP protein aggregates. c–e NSC34 cells were transfected with control or ATG7 siRNA (100 nmole each) and with SOD1-GFP or SOD1^G93A-GFP expression vectors. After incubation for 48 h, the cells were collected. Cell lysates were loaded to a non-reducing gel followed by western blot analysis with indicated antibodies. The data in Fig. 8d were derived from Fig. 8c in which only the density of protein aggregates (excluding the heavy band of the 53 kDa monomer) was quantified. Data in Fig. 8e were derived from Fig. 8c in which the relative levels of ATG7 and LC3 lipidation were analyzed. *p < 0.05; **p < 0.01