Fig. 4
Anti-TLR2 antibody attenuates the LPS + Aβ-induced glycolytic capacity of microglia. Microglia (1 × 105cells/well) were seeded (100 μl/well) on SeaHorse cell culture microplates and incubated in cDMEM ± LPS (100 ng/ml; Enzo Life Sciences, UK). After 4 h, anti-TLR2 antibody (100 nM) was added followed by the addition of Aβ (10 μM; Invitrogen, UK) 30 min later, as described in detail in the “Methods” section. a. The bioenergetic profile of microglia consisting of three baseline measures of ECAR followed by sequential measures following exposure to glucose (10 mM), oligomycin (20 μM) and 2-deoxy-D-glucose (2-DG; 500 mM) is shown. b LPS + Aβ significantly increased mean glycolytic capacity (*p < 0.05), and this was significantly attenuated when cells were also incubated with anti-TLR2 antibody (+p < 0.05; LPS + Aβ vs LPS + Aβ+anti-TLR2 antibody; (F3,13) = 4.55; p = 0.026; one-way ANOVA; Newman-Keuls multiple comparison test). c, d. LPS + Aβ significantly increased the inducible (c) but not the constitutive (d) form of PFKFB3 (*p < 0.05), and anti-TLR2 antibody significantly attenuated the LPS + Aβ-induced change (+p < 0.05; LPS + Aβ vs LPS + Aβ+anti-TLR2 antibody; (F3,18) = 6.067; p = 0.0059 one-way ANOVA, Newman-Keuls multiple comparison test). e LPS + Aβ did not affect PFKFB1 but incubated with LPS + Aβ+anti-TLR2 antibody significantly increased PFKFB1 (+p < 0.05; LPS + Aβ vs LPS + Aβ+anti-TLR2 antibody; (F3,18) = 4.594; p = 0.0167 one-way ANOVA, Newman-Keuls multiple comparison test). Representative immunoblots are shown. For c–e, data are expressed as the percentage of the protein expression in each treatment group vs the control; three independent experiments were performed