Levels of GAD67, beta-III tubulin, and GAPDH in primary neurons. Primary neurons were genetically manipulated with a Cre-lox system to delete apoE4, and lysates were prepared. GAD67 and GAPDH were detected with mouse primary antibodies and IRDye® 800CW Goat anti-Mouse IgG (green bands. Beta-III tubulin was detected with rabbit primary and IRDye 680RD Goat anti-Rabbit IgG (red bands).
Ubiquitination of proteins in N2a cells. Cells were treated with MG132 to induce ubiquitination, and this response was reduced by addition of N-acetyl cysteine (NAC). Inhibition of HSP70 with VER155008 abolished the effect of NAC. Ubiquitinated proteins (green smears) were quantified and normalized against GAPDH as a loading control (red bands). IRDye 800CW goat anti-mouse IgG was used for detection of ubiquitin.