Fig. 1.

PTG overexpression increases glycogen content in cultured astrocytes. Primary cultures of mouse astrocytes were infected with adenovirus containing Flag-tagged mouse PTG (Flag-PTG), or enhanced GFP (EGFP) as control, for five days. (A) mRNA expression levels of genes related to astrocytic glycogen metabolism. Data are means ± SEM and expressed as percentage of the EGFP values for each gene (n = 12–14, 4–5 independent experiments). (B) Representative Western blot of Flag-PTG protein detection (actin as a loading control). Similar results were obtained in 5 independent experiments. (C) Representative immunostaining of Flag-PTG, with (C₁) and without (C₂) nuclear staining with Hoechst. No Flag-PTG staining was observed in the nuclei. Similar results were obtained in 2 independent experiments. (D) Glycogen content. Data are means ± SEM and are expressed as nmoles of glycosyl units per mg of proteins (n = 12, 4 independent experiments). ***P ≤ 0.001 vs respective EGFP conditions with ANOVA followed by Bonferroni's post hoc test (A), or unpaired t-test (D). Pygb, brain glycogen phosphorylase; Gys1, muscle glycogen synthase 1; Gyg, glycogenin; PPP1R6 (or PPP1R3D), Protein phosphatase 1 regulatory subunit 3D; PTG (or PPP1R3C), Protein phosphatase 1 regulatory subunit 3C.