Fig. (1). WISP1 modulates post-translational phosphorylation of AMPK, Akt1, TSC2, and p70S6K during Aβ exposure.

(A) Equal amounts of microglial protein extracts (50 µg/lane) were immunoblotted at 6 hours following Aβ exposure with total and anti–phospho p-AMPK (Thr¹⁷²), p-Akt1 (Ser²⁴⁴⁸), p-TSC2 (Ser¹³⁸⁷/Thr¹⁴⁶²), and p-p70S6K (Thr³⁸⁹) antibodies. WISP1 (10 ng/ml) was applied to microglia cultures 1 hour prior to Aβ exposure (10 µM). During Aβ exposure, phosphorylation of AMPK, TSC2 Ser¹³⁸⁷, TSC2 Thr¹⁴⁶² were increased. In contrast, WISP1 decreased phosphorylation of AMPK and TSC2 Ser¹³⁸⁷ in the presence of Aβ. WISP1 increased phosphorylation of Akt1, TSC2 Thr¹⁴⁶², and p70S6K during Aβ exposure. Representative images for western blot analysis are illustrated. (B) Quantitative analysis of western analysis from 3 experiments was performed using the public domain NIH Image program (US National Institutes of Health at http://rsb.info.nih.gov/nih-image/) (*P < 0.01 vs. Control; ^†P<0.01 vs. Aβ treated alone). Control = untreated microglia. Each data point represents the mean and SD.