Research Paper Volume 13, Issue 13 pp 17097—17117

Figure 13. Drug combinations reduce the expression of amyloid precursor protein (APP) via autophagy. (A) HCT116 ATG16L1 T300 cells were transfected to express wild type APP or mutant APP proteins, APP715 or APP692. In parallel, cells were transfected with a scrambled siRNA control or with siRNA molecules to knock down the expression of Beclin1 or ATG5. After 24h, cells were treated with vehicle control, neratinib (50 nM), AR12 (2 μM) or the drugs in combination for 6h. Cells were fixed in place and the expression of APPs determined by in-cell immuno-staining. (n = 3 +/-SD). * p < 0.05 less than vehicle control; ** p < 0.05 less than neratinib as a single agent. (B) HCT116 ATG16L1 T300 cells were transfected with plasmids to express wild type APP, APP715 or APP692. After 24h, cells were treated with vehicle control, fingolimod (FTY, 100 nM), MMF (5 μM) or the drugs in combination for 6h or 12h. Cells were fixed in place and the expression of the APPs determined by in-cell immuno-staining. (n = 3 +/-SD). * p < 0.05 less than vehicle control; ** p < 0.05 less than MMF alone value. (C) HCT116 ATG16L1 T300 cells were transfected to express wild type APP or mutant APP proteins, APP715 or APP692. In parallel, cells were transfected with a scrambled siRNA control or with siRNA molecules to knock down the expression of Beclin1 or ATG5. After 24h, cells were treated with vehicle control or with [fingolimod, 100 nM plus MMF, 5 μM]. After 6h cells were fixed in place and the expression of the APPs determined by in-cell immuno-staining. (n = 3 +/-SD). * p < 0.05 less than vehicle control.