Fig. 2

LOTUS overexpression suppresses Aβ-induced cofilin dephosphorylation and downregulation of PSD95 expression. a Analysis of functional blocking antibody against PirB. Representative images of the binding assay in Cos7 cells overexpressing PirB. The cells were treated with biotin-conjugated-human Aβ and PirB functional blocking antibodies and immunostained by PirB antibodies. Scale bars, 100 µm. b Quantification of Aβ binding with PirB overexpressed in Cos7 cells. Each binding level of Aβ was normalized to the binding level of Aβ (100 nM) in Cos7 cells overexpressing PirB treated with normal IgG, and the data are presented as mean ± SEM (n = 3, one-way ANOVA post hoc Tukey test *p < 0.05, **p < 0.01). c Representative images of western blotting of the phosphorylated coffin (p-cofilin), cofilin, PSD95, and β-actin in cultured hippocampal neurons of WT mice treated with Aβ and normal IgG or PirB functional blocking antibody. d, e Quantification of normalized p-cofilin and normalized PSD95 in c. Proportion of p-cofilin or PSD95 level was normalized to that treated with PBS and normal IgG-treated samples, and the data are presented as mean ± SEM (n = 5, Two-way ANOVA post hoc Tukey test *p < 0.05, **p < 0.01). f Representative images of immunoblots of the phosphorylated coffin (p-cofilin), cofilin, PSD95, and β-actin in cultured hippocampal neurons of WT or LOTUS-tg mice treated with Aβ. g, h Quantification of normalized p-cofilin and PSD95 in c. The level of p-cofilin or PSD95 was normalized to the level of control treated with PBS in samples of each genotype, and the data are presented as mean ± SEM (n = 5, One-way ANOVA post hoc Tukey–Kramer test *p < 0.05, **p < 0.01)