Fig. 3
Anti-TLR2 antibody decreases IL-1β secretion from microglia. Microglia were prepared as described in the “Methods” section. Cells were harvested for analysis of IL-1β and TNFα mRNA by RT-PCR, or for staining of caspase 1, and supernatant samples were collected and assessed for cytokine concentration. Cell lysate was prepared for analysis of caspase 1 activity using a commercially available kit. a–d LPS + Aβ significantly increased supernatant concentrations of IL-1β (a) and TNFα (c) and also IL-1β mRNA (b) and TNFα mRNA ((a, (F3,14) = 10.87; p = 0.0010; c, (F3,10) = 3.386; p = 0.0745; b, (F3,16) = 11.98; p = 0.0004; d, (F3,18) = 17.10; p < 0.0001 one-way ANOVA **p < 0.01; ***p < 0.001; Newman-Keuls multiple comparison test). Anti-TLR2 antibody significantly attenuated the LPS + Aβ-induced increase in IL-1β but did not affect the changes in IL-1β mRNA, TNFα or TNFα mRNA. e LPS + Aβ significantly increased caspase 1 expression and activity but was not attenuated after anti-TLR2 treatment ((F3,32) = 10.11; p < 0.0001 one-way ANOVA ***p < 0.001). f Caspase 1 activity was increased by LPS + Aβ (*p < 0.05), and anti-TLR2 antibody attenuates this LPS + Aβ-induced effect (+p < 0.05; LPS + Aβ vs LPS + Aβ+anti-TLR2 antibody; ((F3,10) = 4.874; p = 0.0326 one-way ANOVA; Newman-Keuls multiple comparison test). For e and f, data are expressed as the percentage of the caspase 1 expression and activity in each treatment group vs the control; three independent experiments were performed