All reagents were obtained from Sigma-Aldrich (St. Louis, MO, USA), unless otherwise stated.
Intracellular amyloid toxicity in MC65 nerve cells
The MC65 nerve cells were obtained from Dr. L. W. Jin (UC Davis). The induction of intracellular amyloid toxicity in MC65 nerve cells was performed exactly as described8. MC65 nerve cells express the C99 fragment of the amyloid precursor protein (APP) under the control of a tetracycline-mediated promoter. Cells are grown in Dulbecco’s Modified Medium plus 10% fetal calf serum and 2 μg/ml tetracycline. The removal of tetracycline induces C99 synthesis and C99 is cleaved to Aβ. Usually, by day 3 cells in the absence of tetracycline are dead, while control, uninduced cells remain viable. Cell viability was determined by the 3-(4, 5-dimethylthiazolyl-2)-2,5- diphenyltetrazolium bromide (MTT) assay as previously described45. In all cases,…
