Research Paper Volume 13, Issue 13 pp 17097—17117

Figure 10. AR12 and neratinib in combination reduce the expression of TDP-43 via autophagy. (A) HCT116 ATG16L1 T300 cells were transfected to express TDP-43. Twenty-four h later, cells were treated with vehicle control, neratinib (50 nM), AR12 (2 μM), fingolimod (FTY, 100 nM), MMF (5 μM) or the drugs in combination as indicated for 6h or 12h. Cells were fixed in place and the expression of each protein plus ERK2 as a loading control determined by in-cell immuno-staining. (n = 3 +/-SD). * p < 0.05 less than vehicle control. (B) HCT116 ATG16L1 T300 cells were transfected to express TDP-43 and co-transfected with a scrambled siRNA or with siRNA molecules to knock down either Beclin1 or ATG5. Twenty-four h later, cells were treated with vehicle control, neratinib (50 nM), AR12 (2 μM), fingolimod (FTY, 100 nM), MMF (5 μM) or the drugs in combination as indicated for 12h. Cells were fixed in place and the expression of each protein plus total ERK2 as a loading control determined by in-cell immuno-staining. (n = 3 +/-SD). * p < 0.05 less than vehicle control.