Additional file 1 of p47phox deficiency improves cognitive impairment and attenuates tau hyperphosphorylation in mouse models of AD
Creators: name: Gong, Ping
name: Chen, Yan-qing
name: Lin, Ai-hua
name: Zhang, Hai-bo
name: Zhang, Yan
name: Ye, Richard D.
name: Yu, Yang
Publisher: name: figshare
Year: 2020
Subjects: subject: Biochemistry
subject: Cell Biology
subject: Molecular Biology
subject: Neuroscience
subject: Physiology
subject: FOS: Biological sciences
subject: Mental Health
subject: Hematology

description: Additional file 1: Figure S1. The effect of p47phox deficiency on the locomotor, spontaneous exploratory, and anxiety of APP/PS1 mice. The locomotion and spontaneous exploratory activity of mice aged 9 (a, b) and 12 (g, h) months was evaluated on Rotarod and open field tests, respectively. The anxiety of mice aged 9 months (c, d) and 12 months (i, j) was examined in open field and an elevated plus maze. Representative traces of a mouse’s movements during the open field test (e, k) and the elevated plus maze (f, l) are shown. Figure S2. p47phox deficiency does not affect APP levels in APP/PS1 mice. Representative Western blots showing APP expression in the hippocampus and the cortex of WT, APP/PS1, and APP/PS1-Ncf1−/− mice aged 9 months (a) and 12 months (c). b, d Quantification of immunoreactivity of Western blots, normalized against GAPDH. Figure S3. p47phox deficiency attenuates tau hyperphosphorylation in the cortex of APP/PS1 mice and ICV-STZ mice. Representative Western blots showing tau phosphorylation at S199, T205, S396, and S404 in the cortex of WT, APP/PS1, Ncf1−/−, and APP/PS1-Ncf1−/− mice aged 9 months (a) and 12 months (c), and of WT and Ncf1−/− mice receiving ICV injection of STZ or saline at the age of 6 months (e). (b, d, f) Quantifications of the immunoreactivity of Western blots are shown. Figure S4. p47phox deficiency attenuates tau aggregation and NTs in the hippocampus and the cortex of APP/PS1 mice. (a) Representative immunofluorescence staining showing the decreased tau aggregation and NTs in APP/PS1-Ncf1−/− mice aged 12 months, compared with age-matched APP/PS1 mice. Quantification of the fluorescence of AT8 is shown. Figure S5. p47phox deficiency attenuates NP tau in the hippocampus and the cortex of APP/PS1 mice. Representative immunofluorescence staining of AT8 and thioflavin-S (Thio-S) showing the decreased NP tau in APP/PS1-Ncf1−/− mice aged 12 months, compared with age-matched APP/PS1 mice. Figure S6. The expression of NOX subunits (p47phox, p67phox, and NOX2/gp91phox) in APP/PS1 mice. Representative Western blots showing p47phox, p67phox, and gp91phox expression in the brain of WT and APP/PS1 mice at the age of 9 months (a) and 12 months (c). (b, d) Quantifications of the immunoreactivity of Western bolts are shown. Figure S7. p47phox deficiency inhibits the activation of astrocytes in APP/PS1 mice. (a) Representative immunofluorescence staining showing the decreased expression of GFAP in APP/PS1-Ncf1−/− mice aged 9 months and 12 months, compared with age-matched APP/PS1 mice. (b) Quantification of the fluorescence of GFAP is shown. Figure S8. p47phox deficiency inhibits the activation of microglia in APP/PS1 mice. (a) Representative immunofluorescence staining showing the decreased expression of Iba1 in APP/PS1-Ncf1−/− mice aged 12 months, compared with age-matched APP/PS1 mice. (b) Quantification of the fluorescence of Iba1 is shown. Figure S9. p47phox deficiency inhibits high glucose-induced activation of primary astrocytes. Primary cultures of astrocytes from WT and Ncf1−/− newborn mice were treated with DMEM with or without high glucose (75 mM and 150 mM) for 24 h and 48 h. (a) Representative Western blots showing the expression of GFAP. (b, c) Quantification of the immunoreactivity of Western blots is shown.