Supplementary Figure 3: TAT-GluN2BCTM–induced degradation of DAPK1 reduces DAPK1 levels in various subcellular compartments in neuron cultures.
From: Rapid and reversible knockdown of endogenous proteins by peptide-directed lysosomal degradation
Bath applications of TAT-GluN2BCTM (25μM, 1h prior to and during NMDA treatment) significantly decreased DAPK1 in nuclear (left, n=4, p<0.001, F(2,9)=19.139), cytosolic (middle, n=4, p=0.006, F(2,8)=10.417) and mitochondrial (right, n=4, p<0.001, F(2,9)=18.597) subcellular fractions 2h after NMDA treatment and washout, as compared to saline control and NMDA-treated group (grey bar). Cells were collected from at least 3 separate primary cultures. Lysates were collected as a mixture of all treatment samples. One-way ANOVA with Tukey post hoc. * compared to control, Δ compared to NMDA-treated group (grey bar). *p<0.05, ΔΔp<0.01; ***, ΔΔp<0.001bars represent relative mean values±s.e.m. normalized to saline control (arbitrarily set as 1). Full-length blots are available in Supplementary Figure 9.
