Research Paper Volume 13, Issue 13 pp 17097—17117

Figure 8. Neratinib and AR12 interact to reduce the expression of Tau and Tau 301L via autophagy. (A) HCT116 cells were transfected with plasmids to express Tau-GFP or Tau 301L-GFP. After 24h, cells were treated for 3h, 6h or 12h with vehicle control, AR12 (2 μM), neratinib (50 nM) or the drugs in combination. Cells were fixed in place and the detection of Tau levels determined using GFP tag fluorescence (n = 3 +/-SD) * p < 0.05 less than vehicle control; ** p < 0.05 less than neratinib single agent value. (B) HCT116 ATG16L1 T300 cells were transfected to express Tau-GFP or Tau 301L-GFP and in parallel with a scrambled siRNA or with siRNA molecules to knock down the expression of Beclin1 or ATG5. After 24h, cells were treated with vehicle control, AR12 (2 μM), neratinib (50 nM) or the drugs in combination for 6h. Cells were fixed in place and in cell immunostaining performed to determine Tau expression (n = 3 +/-SD) * p < 0.05 less than vehicle control; ** p < 0.05 less than neratinib single agent value.