Fig. 5

Effect of DJ-1 deficiency on the PTEN/PI3K/AKT/GSK3β signaling cascade. a Western blots showing the levels of pAKT T308, pAKT S473, and pGSK-3β S9 in primary astrocytes from WT and DJ-1 knockout mice following a 5- min stimulation with IGF-1 (n = 4 WT, n = 3 DJ-1 knockout, cultures taken from separate animals). Total levels of AKT, GSK-3β, and β-actin are also shown. b-d Quantification of pAKT T308 (b), pAKT S473 (c), and pGSK-3β S9 (d) from the blot shown in (a). pAKT T308 was not detected in unstimulated cells and a Welch’s t-test was used to assess the IGF-1 response (b). Two-way ANOVA followed by Bonferroni’s multiple comparison test was used in (c) and (d). e-g Western blots and quantification for PTEN and pPTEN S380 in primary mouse astrocytes from WT and DJ-1 knockout mice (n = 6 WT, n = 6 DJ-1 knockout, experiment performed using technical replicates). h-i PTEN and pPTEN S380 levels in total brain lysates from 14 to 15-month-old WT and DJ-1 knockout mice determined by immunoblot (n = 5 WT, n = 3 DJ-1 knockout). Unpaired t-tests were used to compare groups shown in (e-i)