Fig. 4
From: The anti-inflammatory Annexin A1 induces the clearance and degradation of the amyloid-β peptide
hrANXA1 treatment increases the expression and activity of Aβ-degrading enzyme neprilysin. a Representative Western blots and quantification of synthetic Aβ1–42 monomers expression in the medium of N2asw cells incubated with synthetic Aβ1–42 (0.1 μM), following 0–48-h treatment with 10-nM hrANXA1 assessed by Western blotting using 6E10 antibody (n = 3). b Representative Western blots and quantification of neprilysin protein expression in N2asw cells treated with 4 μg/ml hrANXA1 and/or the non-selective FPR inhibitor Boc1 (5 μg/ml) for 18 h, normalised to β-actin (n = 10–37 samples). c Quantification of neprilysin activity in N2asw cells treated with 4 μg/ml hrANXA1 and/or the non-selective FPR inhibitor Boc1 (5 μg/ml) for 18 h (n = 2–8 samples). d Representative Western blots and quantification of neprilysin protein expression in SK-N-SK cells treated with 2 μg/ml hrANXA1 and/or selective FPR2 inhibitor WRW4 (5 μM) for 18 h normalised to GAPDH (n = 11–21 samples). e Quantification of MME (neprilysin) mRNA expression by qPCR analysis in SK-N-SK cells treated with 2 μg/ml hrANXA1 for 18 h (n = 5–6 samples). f Representative Western blots and quantification of IDE protein expression in N2asw cells treated with 2–4 μg/ml hrANXA1 for 18 h and normalised to GAPDH. g Representative Western blots and quantification of IDE protein expression in SK-N-SK cells treated with 2 μg/ml hrANXA1 and/or selective FPR2 inhibitor WRW4 (5 μM) for 18 h, normalised to GAPDH (n = 10–18 samples). Values shown in graphs represent the mean value ± SEM and are expressed as a percentage change in comparison to the normalized control or fold change of control. Statistical analysis included two-way ANOVA, one-way ANOVA with Bonferroni’s multiple-comparison post-test, Kruskal-Wallis test with Dunn’s multiple-comparison post-test or unpaired Student’s two-tailed t test as appropriate, *p < 0.05, **p < 0.01, ***p < 0.001