Fig. 7

Astrocyte-specific knock down of SQSTM1 exacerbates Aβ plaque formation and cognitive impairment in APP/PS1 mice. a, Experimental scheme for genetic inhibition of astrocytic autophagy in the hippocampus of APP/PS1 mice: Group 1, APP/PS1 mice with AAV-EF1a-DIO-Control shRNA + AAV-pGFAP-Cre.; Group 2, APP/PS1 mice with AAV-EF1a-DIO-SQSTM1 shRNA + AAV-pGFAP-Cre. b, Double immunostaining for GFAP and Aβ plaque in the brain of APP/PS1 mice with or without SQSTM1 shRNA injection. Scale bars: white, 20 μm; black arrows (x, y, & z in 3D view), 2 μm. c & d, Quantification of mean intensity of GFAP (c) and NeuN (d) immunoreactivity in four groups of mice: a total of 10 ROIs measurement from each group of N = 4 mice e, Quantification of volume of Aβ plaque in two groups of mice: APP/PS1 control, 32 Aβ plaque measurements; APP/PS1 + SQSTM1 shRNA, 54 Aβ plaque measurements from N = 4 mice in each group. f, Quantification of the number of Aβ plaque in two groups of mice: APP/PS1 control; APP/PS1 + SQSTM1 shRNA, 8 ROIs from N = 4 mice in each group. g & h, Discrimination index from behavioral analyses of NOR (g) and NOPR (h) in four groups of mice: WT control, N = 8 mice; WT + SQSTM1 shRNA, N = 4 mice; APP/PS1 control, N = 7 mice; APP/PS1 + SQSTM1 shRNA, N = 4 mice. i, Double immunostaining for phosphorylated (p)-Tau (AT8: Ser202/Thr205) and GFAP in the hippocampus of LC3B shRNA or SQSTM1 shRNA virus-injected APP/PS1 mice. Scale bars (white): 20 μm. j, Quantification of p-Tau (Ser202/Thr205) intensity in neurons of DG in three groups of mice: a total of 20 ROIs measurements from N = 3 mice in each group. k, Correlation analysis between GFAP and p-Tau (Ser202/Thr205) intensities (derived from panel j). Data are presented as mean ± SEM. Significantly different at *, p < 0.05; **, p < 0.01