Fig. 4

CCL3 release by astrocytes is ASC dependent and is critical for phagocytosis efficiency. Astrocytes primed with 1 μg/ml LPS for 3 h were treated with 10 μM Aβ42 for 3 h. The concentration of CCL3 in culture supernatants were then measured by ECLIA (a) (Control n = 4 in duplicate; LPS Aβ42 n = 7 in duplicate). Astrocytes were treated with 0.1 μg/ml of CCL3-neutralizing antibody 15 min before Aβ42 and phagocytic capacity was assessed by adding pHrodo™ Red Zymosan Bioparticles® 2 h before visualization (b, c). Primary astrocytes were treated with increasing concentrations of mouse recombinant CCL3 for 2 h during exposure to bioparticles. The amount of internalized bioparticles was evaluated by measuring fluorescence areas using image J (c, n = 4 in duplicate; e, n = 3 in triplicate). Results are expressed as a percentage of total area and are the mean ± SEM, and statistical differences were determined using an ANOVA analysis and post Bonferroni multiple comparison test: ***P < 0.001, **P < 0.01, and *P < 0.05 vs respective controls (a, e), ⁺⁺⁺ P < 0.001 and ⁺⁺ P < 0.01 (a); **P < 0.01 (c)