Figure 4

GSK3β-positive inclusions in conditional Atg7 -deficient mice. (a-c) GSK3β-positive inclusions in cortical neurons of CamK-Atg7 cKO mice. a, An antibody recognizing total GSK3β (red, arrows), stained p62-positive inclusions (green) in cortical neurons of CamK-Atg7 cKO mice. Bar, 10 μm. b, Antibodies recognizing phosphorylated, activated form of GSK3β (Tyr279/Tyr216; in red, arrows) stained p62-positive (green) inclusions in cortical neurons of CamK-Atg7 cKO mice. Bar, 10 μm. c, An antibody recognizing phosphorylated inactivated forms of GSK3β at Ser9 residues (red, arrows), stained p62-positive inclusions (green) in cortical neurons of CamK-Atg7 cKO mice. Bar, 10 μm. (d) GSK3β levels are elevated in the forebrain extracts of CamK-Atg7 cKO mouse. Western blotting reveals that phosphorylated forms of GSK3β (both activated Tyr216 residue and inactivated Ser9 residue) as well as total GSK3β were significantly increased in CamK-Atg7 cKO brain tissue extracts. Triangle, GSK3α. Circle, GSK3β. n = 5 per group. (e) Summary of phosphorylation and antibody recognition sites of human tau protein. The phosphorylated sites (Ser202/Thr205 [AT8], Thr212/Ser214 [AT100], and Thr231/Ser235 [TG3] residues) in Atg7-deficient mice are shown in red. Non-phosphorylated sites are shown in blue. Phosphorylation sites are numbered according to the human tau protein by convention; the homologous corresponding phosphorylation sites in the mouse tau protein are each positioned 11 amino acids towards the amino terminus.