Skip to main content
. 2022 Mar 3;16:801179. doi: 10.3389/fncel.2022.801179

FIGURE 2.

FIGURE 2

Characterization of second-generation CRBN-recruiting degraders (FMF-06-series) focusing on optimization of linker attachment chemistry. (A–C) The FMF-06-series was synthesized based on the T807 core scaffold for tau recognition (A), thalidomide as the E3 ligand for CRBN engagement (B), and a variable linker length and attachment composition (C). (D) Summary of chemical properties of the FMF-06-Series. (E–J) Degraders’ concentration effect on tau protein of A152T neurons (6-week differentiated) with analysis of total tau (TAU5) and P-tau S396 levels upon treatment for 24 h. Representative western blots are shown (E) and brackets indicate bands quantified for TAU5 and P-tau S396 densitometry (F–J). Data points represent mean densitometry normalized to actin and relative to vehicle ± SEM (n = 3). (K–P) Degraders’ concentration effect on tau protein levels of A152T neurons (6-week differentiated) after a 4 h treatment. Representative western blots are shown (K) for immunoprobing with TAU5, P-tau S396 and actin. Data points (L–P) represent mean densitometry normalized to actin and relative to vehicle ± SEM (n = 3). The 0 μM degrader data points correspond to vehicle alone (DMSO) control treatment. Statistics: 2-tailed unpaired Student’s t-test for each concentration relative to vehicle; nsP > 0.05, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.