Fig. 3
CLIC1 knockdown in microglia ameliorates cognitive function, regulates microglial polarization and ameliorates AD pathology. (A, B) CLIC1 expression in the hippocampus of 6-month-old (A) and 10-month-old (B) APP/PS1 and age-matched WT mice was detected using RT-PCR assay, n = 4. (C) RT-PCR assay was conducted to detect the expression of CLIC1 in the hippocampus of mice after hippocampal microglia were infected with shCLIC1 AAVs for 6 weeks, n = 4. (D ~ F) The effect of CLIC1 knockdown on the cognitive function of APP/PS1 mice was evaluated using MWM test (D), passive avoidance test (E) and novel object recognition task (F), n = 8 ~ 10 mice. (G) CLIC1 expression was detected using RT-PCR assay after BV-2 cells were transfected with shCLIC1 plasmids, n = 3. (H ~ M) The effects of CLIC1 knockdown on the expression of Iba-1, TNF-α, Pro- and cleaved IL-1β, CD16, Arg1 and IL-10 in Aβ-treated BV-2 cells were assessed using Western blot assay, n = 4. (N ~ U) The effects of CLIC1 knockdown on the expression of Iba-1, TNF-α, Pro- and cleaved IL-1β, CD16, Arg1, IL-10, Aβ42 and p-tau in the hippocampus of APP/PS1 mice were evaluated using Western blot assay, n = 3 ~ 4. (V) The effect of CLIC1 knockdown on Aβ phagocytosis of BV-2 cells was assessed using flow cytometry assay, n = 3 ~ 4. (W) Golgi staining assay was performed to assess the effect of CLIC1 knockdown on spine density in the hippocampus of APP/PS1 mice, n = 3 mice. All data in the figure are presented as mean ± SEM. Student’s t test (A ~ G, O ~ W) and one-way ANOVA (I ~ M) were used to assess statistically significant differences. *P < 0.05, **P < 0.01