Figure 5:

Impact of T-Loop and CPE Cys-to-Ala mutations on BRSK redox sensitivity in a cellular EGFP-Tau HEK-293T co-expression system. Representative immunoblot of EGFP-Tau co-expressed with WT and Cys-to-Ala mutants of (a) BRSK1 and (b) BRSK2 (left panels). Transiently transfected HEK-293T cells were treated with or without 10 mM H₂O₂ for 10 mins before the addition of 20 mM GSH. Whole cell lysates were harvested after a further 15 mins. Signal density for phospho Tau S262 and total Tau (GFP) was obtained using ImageStudio software (Licor) and results from at least 3 biological replicates were anaylzed with Graphpad Prism software using one way anova to determine significance (right panels). Data shown is mean and SE. All values are normalized to Tau pS262 signals from control (buffer only treatment) WT BRSK and Tau co-transfections. Data shown is mean and SD. * = P < 0.05, **= P < 0.01, ***= P < 0.001.