Sf1670

SF1670 (B-0350, Echelon Biosciences), dissolved in dimethylsulfoxide (DMSO), was added to the culture medium 24 h after cell seeding. DMSO was used as the vehicle control. After 24 h of treatment, the cells were fixed for immunostaining and imaging.

Microglia and M‐MØP cells were prepared as described above. Cells were then exposed to anti‐FcγRII/III (2.4G2) antibody at 5 µg/ml or oligomers of Aβ1–42 (40 µM). Designated wells were inhibited by pre‐exposure for 2 h with 3‐a‐aminocholestane (20 µM, B‐0341), LY294002 (10 µM, B‐0294) or SF1670 (5 µM, B‐0350) (Echelon Biosciences). Cells were then exposed to ice‐cold 0.5 M TCA (trichloroacetic acid T6399, Sigma) for 5 min. Cells were then scraped off and spun (1,720 g, 7 min, 4°C). The cell pellet was washed twice with 5% TCA/ 1 mM EDTA, vortexed for 30 s and centrifuged (1,720 g, 5 min). The pellet is then vortexed twice with MeOH: CHCl3 (2:1) for 10 min at room temperature and centrifuged (1,720 g, 5 min). The pellet is then vortexed with MeOH: CHCl3:12 N HCl (80:40:1) for 25 min at room temperature and centrifuged at 1,720 g for 5 min. The pellet is then discarded, and CHCl3 and 0.1 N HCl are added. The tube is then centrifuged at 1,720 g for 5 min to separate organic and aqueous phases. Collect the organic phase into a new vial and dry in a vacuum dryer for 60 min.