Fig. 5.
FoxG1 inhibits the Aβ-induced activation of NLRP3 inflammasome via the upregulation of autophagy in N2A cells. A FoxG1/N2A cells and pCDH/N2A cells are transiently transfected with APPswe plasmid for 48 h. The expressions of Aβ, NLRP3, TNF-α and IL-6 were studied by western blot. (B: Aβ [F2,15 = 367.3, P < 0.0001], NLRP3 [F2,15 = 82.98, P < 0.0001], TNF-α [F2,15 = 79.66, P < 0.0001], IL-6 [F2,15 = 54.51, P = 0.0001]). Bar chart indicates the relative expression of these proteins in (A). C N2A cells transiently co-transfected with siRNA control or siRNA-FoxG1 and APPswe plasmid for 48 h; the expressions of NLRP3, TNF-α, and IL-6 were studied by western blot. (D: NLRP3 [t10 = 7.655, P = 0.0016], TNF-α [t10 = 11.80, P = 0.0003], IL-6 [t10 = 11.58, P = 0.0003]). Bar chart indicates the relative expression of these proteins in (C). E FoxG1/N2A cells and pCDH/N2A cells are transiently transfected with APPswe plasmid for 48 h, the expressions of LC3, Beclin1, SQSTM1/P62 and AMPK, p-AMPK, mTOR, and p-mTOR proteins were studied by western blot. (F Beclin1 [F2,15 = 33.89, P = 0.0005], SQSTM1/P62 [F2,15 = 67.36, P < 0.0001]), Bar chart indicates the relative expression of Beclin1 and SQSTM1/P62 in (E). (H [F2,15 = 10.19, P = 0.0118], I [F2,15 = 13.76, P = 0.0057]). Bar chart indicates the expression of p-AMPK/AMPK or p-mTOR/mTOR in (E). (G [F2,15 = 8.809, P < 0.0076]), Bar chart indicates the expression of LC3 II/LC3 I in (E). J FoxG1/N2A cells and pCDH/N2A cells transiently transfected with APPswe plasmid were treated with 10 mM 3-MA for 48 h or no 3-MA treatment. After 48 h, cells were then harvested for the protein detections of NLRP3, TNF-α, and IL-6 by western blot. (K [F5,30 = 21.39, P < 0.0001], L [F5,30 = 25.39, P < 0.0001], M [F5,30 = 34.61, P < 0.0001]). Bar chart indicates the relative expression of these proteins in (J). GAPDH or Tubulin was used as loading control. Values are expressed as means ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001. For each group, n = 6/group