1. Fig. 8 C9orf72 knockdown results in an increase in autophagic flux.
C9orf72 knockdown results in an increase in autophagic flux. (A) Confocal imaging of GFP-mCherry expression in N2A cells infected with lentiviruses expressing control or C9orf72 shRNA. (B) Quantification of the percentage of yellow dots, which maintains both GFP and mCherry signal, out of total red dots. Values represent SEM of three independent experiments; ~50 cells were randomly selected for each statistical analysis. *P < 0.05 (Student’s t test). (C) Western blot analyses of p62 expression. Wild-type or Smcr8 mutant MEFs were infected with lentiviruses expressing control or C9orf72 shRNA followed by 72 hours of culture before protein lysate collection. β-Actin serves as the loading control. (D) Quantification of p62 levels relative to actin. Error bars represent SEM of three measurements from three independent experiments; *P < 0.05 (Student’s t test). n.s. represents no significant difference detected. (E and G) Western blot analysis of cathepsin D (E) or cathepsin L (G) expression in the control or C9orf72 knockdown MEFs. β-Actin serves as the loading control. (F and H) Quantification of cathepsins D or L expression. Error bars represent SEM of three measurements from three independent experiments; *P < 0.05, ***P < (Student’s t test); n.s. represents no significant difference detected.
Mei Yang et al. Sci Adv 2016;2:e
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