Fig. 1
A-SMase is activated downstream of the TLR4 receptor and translocated to the plasma membrane in response to Aβ oligomers. A A-SMase activity expressed as percentage of control measured in microglia treated with Aβ1−42 oligomers in the presence of Imipramine (10 µM) or TNFα receptor inhibitor (10 ng/mL) or TLR4 receptor inhibitor (10 µM). Bar graphs represent average ± SD of N > 3/group One-way ANOVA, Dunnett’s posthoc testing (**p < 0.01, ***p < 0.001, ****p < 0.0001). B Photomicrographs of primary microglia stimulated with or without Aβ1−42 peptide conjugated to Alexa 555 (Red) and subsequently immunolabeled with anti-Cer (Green) and anti-A-SMase (Magenta) antibodies. Nuclei were stained with DAPI. Scale bar = 20µm. C Violin plots depicting Pearson correlation coefficient between Cer and A-SMase fluorescent intensities of 4–5 photomicrographs / condition. Unpaired t-test (**p < 0.01. D IL-1α, TNFα and C1q levels measured in supernatant of primary microglia by ELISA after 18 h incubation with Aβ1-42 oligomers and of 10 µM Imipramine or 5 µM ARC39. Bar graphs represent average ± SD of N > 3/group. One-way ANOVA, Dunnett’s posthoc testing (*p < 0.05, **p < 0.01, ****p < 0.0001)