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. 2017 Mar 8;37(10):2709–2722. doi: 10.1523/JNEUROSCI.4559-15.2017

Figure 5.

Figure 5.

ARPP-16/19 regulates PP2A in a substrate-specific manner in striatum. a, Samples from different brain regions (as indicated) from WT and conditional ARPP-16/19 knock-out (cKO) mice were analyzed by immunoblotting with an antibody that recognizes ARPP-16, ARPP-19, and ENSA. Striatal slices from ARPP-16/19 cKOs (n = 4) or WT littermate controls (n = 3 or 4) were isolated and basal phosphorylation of the PP2A targets, (b) DARPP-32 at Thr75, or (c) Akt at Thr308 were analyzed by SDS-PAGE and immunoblotting. b, Phospho-Thr75 (pD32–T75) and total DARPP-32 (totD32) blots. c, Phospho-Thr308 (pAkt-t308) and total Akt (totAkt) blots. Bottom bar graphs represent cumulative data. *p < 0.05 (Student's t test). Error bars indicate SEM. d, Striatal slices from ARPP-16/19 cKOs (n = 4) or WT littermate controls (n = 4) were incubated with 10 μm forskolin for 0, 10, 30, or 60 min. Samples were analyzed by SDS-PAGE, and immunoblotting was done for phospho-Thr202/204 and total ERK1/2 (top); quantitation of phosphorylation as done for Thr202/204 normalized to ERK1. Bottom graph represents cumulative data. **p < 0.01 (multivariate ANOVA). b–d, Phospho-site signals were each normalized to total protein, and then data for cKO mouse samples were normalized to controls.