Supplementary Figure 1: Design of the CMA targeting peptide and time-dependent degradation by CMA.
From: Rapid and reversible knockdown of endogenous proteins by peptide-directed lysosomal degradation
(a) Schematic diagram illustrating the design of the targeting peptide mediated protein degradation. Following administration, the targeting peptide enters the cell through its cell membrane penetrating domain (CMPD), binds to the target protein via its protein binding domain (PBD), and chaperones the peptide-protein complex to the lysosome for degradation via its chaperone-mediated autophagy targeting motif (CTM). (b) CTM-GFP is degraded in a time-dependent manner, without affecting the stability of endogenous CMA-substrate GAPDH in HEK cells transiently transfected with CTM-GFP. N=3 from 3 separate cultures and transfections. One-way ANOVA F(9,20)=12.418, p<0.001. Bars represent protein levels normalized to 12h WT-GFP (white bar), and compared to 12h WT-GFP (white bar, *) or 12h CTM-GFP (grey bar, Δ) *,Δ p<0.05, **,ΔΔ p<0.01 and ***, ΔΔΔp<0.001, bars represent relative mean values±s.e.m. Full-length blots are available in Supplementary Figure 9.
