Fig. 6

Rapamycin alleviates p62 accumulation in cells treated with patient iPSC-derived ACM. HEK293T cells were treated with control (n = 3) and patient (n = 3) ACM for 5 days, with either 200nM Rapamycin or 100nM Trehalose for the final 2 days where indicated. Cells were then fixed in 4% paraformaldehyde and stained for p62 (a), or protein harvested to assess changes in SOD1 expression (b). P62 puncta were analyzed using automated image analysis. Trehalose increased p62 puncta in HEK293T cells treated with control ACM, but not patient ACM. Rapamycin decreased levels of p62 puncta in HEK293T cells treated with patient ACM, but had no effect on cells treated with control ACM. Rapamycin had no effect on SOD1 levels following treatment with ACM. *p < 0.05 and **p < 0.01 were considered statistically significant. Error bars represent standard error of mean