Supplementary Figure 6: Immunoblots showing that the macroautophagy inhibitor 3-methyladenine (3-MA) cannot rescue targeting-peptide mediated degradation.
From: Rapid and reversible knockdown of endogenous proteins by peptide-directed lysosomal degradation
(a) α-synuclein targeting peptide TAT-βsynCTM (25μM) significantly decreased native α-synuclein in the presence of 3-methyladenine (10mM). Basal level of α-synuclein showed a non-significant trend of increase with the addition of 3-methyladenine. n=5, p<0.001, F(3,16)=47.013. (b) 3-methyladenine did not alter either the basal levels or NMDA activation-dependent knockdown of DAPK1 by its targeting peptide TAT-GluN2BCTM. n=4, p<0.001, F(3,12)=21.675. (c) Similarly, 3-methyladenine failed to affect either the basal or the TAT-GluN2B9cCTM reduced level of PSD-95. n=4, p<0.001, F(3,12)=14.836. β-actin was used as loading control. One-way ANOVA with Tukey post hoc. * p<0.05, **p<0.01 and ***p<0.001; bars represent relative mean values±s.e.m. normalized to saline control (arbitrarily set as 1). Cells were collected from at least 3 separate primary cultures. Full-length blots are available in Supplementary Figure 9.
