Effect of cholesterol on the cleavage efficiency of purified
γ-secretase in detergent-free proteoliposomes. A, the
effect of various concentrations of cholesterol on Aβ40 production in
proteoliposomes of optimal phospholipid or sphingolipid compositions from
Fig. 3 (PC(90)/PE(10),
PC(90)/PS(10), PC(90)/PA(10), PC(90)/SM(10), PC(95)/CS(5), PC(90)/GS(10)) was
examined. Error bars represent mean ± S.E. Data were
statistically analyzed using the unpaired, two-tailed Student's t
test with Bonferroni correction. Statistical significance of p <
0.05 (*), p < 0.01 (**), p <
0.001 (***), and p < 0.0001 (****)
versus the PC/PE-detergent-containing control is indicated.
B, the effect of various concentrations of cholesterol on Aβ42
production in proteoliposomes (PC(90)/PE(10) and PC(90)/GS(10)) was examined.
Error bars represent mean ± S.E. Data were statistically
analyzed using the unpaired, two-tailed Student's t test with
Bonferroni correction. Statistical significance of p < 0.05
(*), p < 0.001 (**), and p <
0.0001 (***) versus the 0% cholesterol control is
indicated. C, AICD levels of samples from the C100FLAG assay in
A and B were analyzed by Western blot. AICD fragments were
detected with the M2 FLAG antibody. D, the effect of various
concentrations of cholesterol on NICD production in proteoliposomes of PC/PE
in detergent, PC(100), PC(90)/PE(10)/0.0063% cholesterol, and
PC(90)/GS(10)/0.013% cholesterol was examined. NICD levels were analyzed by
Western blot and the NICD was detected with the 1744 antibody (Cell Signaling
Technologies).