Thiamet-G treatment of APPSwe-expressing 20E2 cells and primary hippocampal neurons does not alter APP processing. A. 20E2 cells were treated with Thiamet-G for 16 hours, and then the amount of Aβ42 in the cell culture media, collected at the indicated time points after the replacement of new culture media, was assessed using an ELISA assay and corrected for the amount of protein in the cell lysates from each condition. This analysis reveals that Thiamet-G does not alter the amount of Aβ42 that is produced. B. Little Aβ42 is detected in untransfected human embryonic kidney (HEK) cells indicating that validity of this culture model. C. O-GlcNAc levels in the 20E2 cells are increased at all time points analyzed (*indicates p <0.05, paired two-tailed t-test). D. Levels of APP-CTF were unchanged in 20E2 cells treated with Thiamet-G for 24 hours. E. Aβ42 production is not altered in primary hippocampal neurons treated with Thiamet-G for 24 hour. F. APP-CTF levels are unchanged in 24 hour Thiamet-G treated hippocampal neurons. G. O-GlcNAc levels in primary hippocampal neurons are increased after 24 hours of treatment with Thiamet-G. (***indicates p <0.001,, paired two-tailed t-test) Error bars represent standard error of the mean (± S.E.M). N.S. Not Significant. For all panels, N =3.