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. Author manuscript; available in PMC: 2019 May 23.
Published in final edited form as: Biochem Biophys Res Commun. 2018 Apr 9;499(4):751–757. doi: 10.1016/j.bbrc.2018.03.209

Fig.1. Cambinol inhibits nSMase2 activity in cell extracts and EV-mediated tau seed propagation.

Fig.1

(A) Schematic representation of the “D+R” and EV-mediated assays. P2 - synaptosomal fraction from human AD brain. (B) Imaging flow cytometry: top row - DID-labeled recipient cell, cultured separate from donor cells; middle row - seeded donor cells with FRET-positive tau inclusions; bottom row - DID-labeled recipient cells with FRET-positive tau inclusions after co-culturing with seeded donor cells. (C) Dose-dependent reduction in the tau transfer from donor to recipient cells after treatment with cambinol (D+R assay). (D) Dose-dependent inhibition of nSMase2 activity by cambinol. (E) Treatment of P2-seeded donor cells with cambinol (P2/Camb) or GW4869 (P2/GW4869) at 50μM shows decrease in EV-mediated transfer of tau to recipient cells. Lipo/DMSO group - EVs derived from vehicle treated donor cells treated with empty liposomes and DMSO), P2/DMSO –EVs derived from P2-seeded donor cells treated with DMSO. * p<0.05, ** p<0.01, *** p<0.01.